A new pterocarpan glycoside, （-）-maackiain 3-O-6＇-O-acrylyl-β-D-galactopyranoside（1） was isolated from the whole parts of Caragana jubata（pall.） Poir., together with three known pterocarpan glycosides, （-）-m...A new pterocarpan glycoside, （-）-maackiain 3-O-6＇-O-acrylyl-β-D-galactopyranoside（1） was isolated from the whole parts of Caragana jubata（pall.） Poir., together with three known pterocarpan glycosides, （-）-maackiain 3-O-6＇-O-acetyl-β-D-glucopyranoside（2）, （-）-maackiain 3-O-β-D-glucopyranoside（3） and （-）-maackiain 3-O-β-D-galactopyranoside（4）. All the compounds were isolated from the title plant for the first time. The structure of the new compound was established on the basis of detailed 1D, 2D NMR and circular dichroism spectroscopic analyses. All the compounds were evaluated for their cytotoxic activities against three tumor cell lines, A549, HL-60 and P388.展开更多
During the screening of a traditional Chinese folk herb library against Hep G2 and Hep3 B cell lines, the Et OAc extract from the Tibetan medicine, Caragana tibetica（CT-Et OAc） exhibited potential anti-hepatocellula...During the screening of a traditional Chinese folk herb library against Hep G2 and Hep3 B cell lines, the Et OAc extract from the Tibetan medicine, Caragana tibetica（CT-Et OAc） exhibited potential anti-hepatocellular carcinoma（anti-HCC） activity. HPLC-based activity profiling was performed for targeted identification of anti-HCC activity from CT-Et OAc by MS-directed purification method. CT-Et OAc was separated by time-based fractionation for further anti-HCC bioassay by a semipreparative HPLC column（150 mm × 10 mm i.d., 5 μm） with a single injection of 5 mg. Bioassay-guided and ESIMS-directed large scale purification was performed with a single injection of 400 mg of CT-Et OAc by peak-based fractionation. A 1.4-mm heavy wall micro NMR tube with z-gradient was used to measure one and two dimensional NMR spectra for the minor or trace amounts of components of the extract. Two active compounds could be elucidated as naringenin chalcone（CT-1） and 3-hydroxy-8, 9-dimethoxypterocarpan（CT-2） relevant to anti-HCC effects for the Et OAc extract of C. tibetica rapidly and unambiguously by this protocol.展开更多
Astrapterocarpan(AP) is a bioactive constituent of Astragali Radix and was selected as a model compound for investigating the in vitro metabolism of pterocarpans in this study. Its in vitro metabolism was conducted by...Astrapterocarpan(AP) is a bioactive constituent of Astragali Radix and was selected as a model compound for investigating the in vitro metabolism of pterocarpans in this study. Its in vitro metabolism was conducted by incubation with rat hepatic 9000 g supernatant(S9) in the presence of an NADPH-generating system. At first, four compounds were isolated and their structures were elucidated as 6 a-hydroxy-AP(M1), astrametabolin I [M2, 1 a-hydroxy-9, 10-dimethoxy-pterocarp-1(2), 4-diene-3-one], 9-demethyl-AP(M3, nissolin) and 4-methoxy-astraisoflavan(M4, 7, 2’-dihydroxy-4, 3’, 4’-trimethoxy-isoflavan) on the basis of NMR data, respectively. Among them, M1, M2 and M4 were new compounds. Next, the metabolite profile of AP in rat hepatic S9 was obtained via HPLC-DAD-ESI-IT-TOF-MSn, and 40 new metabolites were tentatively identified. These newly identified metabolites included 9 monohydroxylated metabolites, 1 demethylated metabolite, 7 demethylated and monohydroxylated metabolites, 4 dihydroxylated metabolites, 1 hydration metabolite, 1 didemethylated metabolite, 2 glucosylated metabolites, 1 monohydroxylated and dehydrogenated metabolite, 2 monohydroxylated and demethylated and dehydrogenated metabolites, 2 dimerized metabolites, 3 dimerized and monohydroxylated metabolites, 2 dimerized and didemethylated metabolites, and 5 dimerized and demethylated metabolites. Finally, the major metabolic reactions of AP in rat hepatic S9 were summarized and found to be hydroxylation, demethylation, dimerization, hydration, and dehydrogenation. More importantly, the biotransformation from AP to M2 and the dimerization of AP by incubation with hepatic S9 were reported for the first time. In conclusion, this is the first report on the metabolism of a pure pterocarpan in animal tissues, and these findings will provide a solid basis for further studies on the metabolism of other pterocarpans.展开更多
Medicarpin is an important bioactive compound with multiple medicinal activities,including anti-tumor,anti-osteoporosis,and anti-bacterial effects.Medicarpin is associated with pterocarpans derived from medicinal plan...Medicarpin is an important bioactive compound with multiple medicinal activities,including anti-tumor,anti-osteoporosis,and anti-bacterial effects.Medicarpin is associated with pterocarpans derived from medicinal plants,such as Sophora japonica,Glycyrrhiza uralensis Fisch.,and Glycyrrhiza glabra L.However,these medicinal plants contain only low amounts of medicarpin.Moreover,the planting area for medicarpin-producing plants is limited;consequently,the current medicarpin supply cannot meet the high demands of medicinal markets.In this study,eight key genes involved in medicarpin biosynthesis were identified using comparative transcriptome and bioinformatic analyses.In vitro and in vivo enzymatic reaction confirmed the catalytic functions of candidate enzymes responsible for the biosynthesis of medicarpin and medicarpin intermediates.Further engineering of these genes in Saccharomyces cerevisiae achieved the heterologous biosynthesis of medicarpin using liquiritigenin as a substrate,with a final medicarpin yield of 0.82±0.18 mg/L.By increasing the gene copy numbers of vestitone reductase(VR)and pterocarpan synthase(PTS),the final medicarpin yield was increased to 2.05±0.72 mg/L.This study provides a solid foundation for the economic and sustainable production of medicarpin through a synthetic biology strategy.展开更多
Main observation and conclusion Five novel pterocarpan derivatives(1—5)and three known pterocarpans were isolated from the roots of Sophora flavescens Ait.Their structures were elucidated based on extensive spectrosc...Main observation and conclusion Five novel pterocarpan derivatives(1—5)and three known pterocarpans were isolated from the roots of Sophora flavescens Ait.Their structures were elucidated based on extensive spectroscopic data(UV,IR,1D and 2D NMR,and HR-ESI-MS).Notably,compounds 1—5 were the first reported natural products,which possess unique pterocarpan-glucose-long chain aliphatic acid structures.The kinds of aliphatic acids were determined by comparison with authentic aliphatic acids using HPLC-ELSD analysis after acid hydrolysis.The absolute configurations of 1—5 were confirmed by analysis of their ECD spectra.All compounds were evaluated against lipopolysaccharide(LPS)-induced TNF-αproduction in RAW264.7 cells,and compound 1 showed significant inhibitory activity with inhibition rate of 86.22%,IC_(50)=0.79μmol/L(dexamethasone as positive control,inhibition rate of 71.38%).展开更多
基金Supported by the National Natural Science Foundation of China(No.30660221)the Natural Science Foundation of Qinghai Province,China(No.2006-N-554)
文摘A new pterocarpan glycoside, （-）-maackiain 3-O-6＇-O-acrylyl-β-D-galactopyranoside（1） was isolated from the whole parts of Caragana jubata（pall.） Poir., together with three known pterocarpan glycosides, （-）-maackiain 3-O-6＇-O-acetyl-β-D-glucopyranoside（2）, （-）-maackiain 3-O-β-D-glucopyranoside（3） and （-）-maackiain 3-O-β-D-galactopyranoside（4）. All the compounds were isolated from the title plant for the first time. The structure of the new compound was established on the basis of detailed 1D, 2D NMR and circular dichroism spectroscopic analyses. All the compounds were evaluated for their cytotoxic activities against three tumor cell lines, A549, HL-60 and P388.
基金supported by grants from Natural Science Foundation of Qinghai Province(No.2012-z-904)the Major Research Project of Department of Education of Hubei Province,China(No.D20131103)+1 种基金National Natural Science Foundation of China(No.81102798)Wuhan Youth Chenguang Program of Science and Technology(No.2013070104010028)
文摘During the screening of a traditional Chinese folk herb library against Hep G2 and Hep3 B cell lines, the Et OAc extract from the Tibetan medicine, Caragana tibetica（CT-Et OAc） exhibited potential anti-hepatocellular carcinoma（anti-HCC） activity. HPLC-based activity profiling was performed for targeted identification of anti-HCC activity from CT-Et OAc by MS-directed purification method. CT-Et OAc was separated by time-based fractionation for further anti-HCC bioassay by a semipreparative HPLC column（150 mm × 10 mm i.d., 5 μm） with a single injection of 5 mg. Bioassay-guided and ESIMS-directed large scale purification was performed with a single injection of 400 mg of CT-Et OAc by peak-based fractionation. A 1.4-mm heavy wall micro NMR tube with z-gradient was used to measure one and two dimensional NMR spectra for the minor or trace amounts of components of the extract. Two active compounds could be elucidated as naringenin chalcone（CT-1） and 3-hydroxy-8, 9-dimethoxypterocarpan（CT-2） relevant to anti-HCC effects for the Et OAc extract of C. tibetica rapidly and unambiguously by this protocol.
基金supported by the National Natural Science Foundation of China(No.81673595)China Postdoctoral Science Foundation(Nos.20080430293 and 200902040)Guizhou Natural Science Foundation(No.QIANKEHE [2018]1071)
文摘Astrapterocarpan(AP) is a bioactive constituent of Astragali Radix and was selected as a model compound for investigating the in vitro metabolism of pterocarpans in this study. Its in vitro metabolism was conducted by incubation with rat hepatic 9000 g supernatant(S9) in the presence of an NADPH-generating system. At first, four compounds were isolated and their structures were elucidated as 6 a-hydroxy-AP(M1), astrametabolin I [M2, 1 a-hydroxy-9, 10-dimethoxy-pterocarp-1(2), 4-diene-3-one], 9-demethyl-AP(M3, nissolin) and 4-methoxy-astraisoflavan(M4, 7, 2’-dihydroxy-4, 3’, 4’-trimethoxy-isoflavan) on the basis of NMR data, respectively. Among them, M1, M2 and M4 were new compounds. Next, the metabolite profile of AP in rat hepatic S9 was obtained via HPLC-DAD-ESI-IT-TOF-MSn, and 40 new metabolites were tentatively identified. These newly identified metabolites included 9 monohydroxylated metabolites, 1 demethylated metabolite, 7 demethylated and monohydroxylated metabolites, 4 dihydroxylated metabolites, 1 hydration metabolite, 1 didemethylated metabolite, 2 glucosylated metabolites, 1 monohydroxylated and dehydrogenated metabolite, 2 monohydroxylated and demethylated and dehydrogenated metabolites, 2 dimerized metabolites, 3 dimerized and monohydroxylated metabolites, 2 dimerized and didemethylated metabolites, and 5 dimerized and demethylated metabolites. Finally, the major metabolic reactions of AP in rat hepatic S9 were summarized and found to be hydroxylation, demethylation, dimerization, hydration, and dehydrogenation. More importantly, the biotransformation from AP to M2 and the dimerization of AP by incubation with hepatic S9 were reported for the first time. In conclusion, this is the first report on the metabolism of a pure pterocarpan in animal tissues, and these findings will provide a solid basis for further studies on the metabolism of other pterocarpans.
基金supported by the National Natural Science Foundation of China(81903526,81991523,82072240,and 32270192)the Open Project of Chinese Materia Medica First-Class Discipline of Nanjing University of Chinese Medicine(No.2020YLXK008 to W.L.)+3 种基金the Open Project of State Key Laboratory of Drug Research,Shanghai Institute of Materia Medica,Chinese Academy of Sciences(No.SIMM2205KF to W.L.)the Open Project of State Key Laboratory of Microbial Resources,Institute of Microbiology,Chinese Academy of Sciences(No.SKLMR-20220704 to W.L.)Jiangsu Graduate Research Innovation Program Project(KYCX22_2024 to CJ.L.)the Open Funding Project of the State Key Laboratory of Bioreactor Engineering,East China University of Science and Technology to W.L.,the Fok Ying Tung Education Foundation,and Jiangsu Specially-Appointed Professor Talent Program to W.L.
文摘Medicarpin is an important bioactive compound with multiple medicinal activities,including anti-tumor,anti-osteoporosis,and anti-bacterial effects.Medicarpin is associated with pterocarpans derived from medicinal plants,such as Sophora japonica,Glycyrrhiza uralensis Fisch.,and Glycyrrhiza glabra L.However,these medicinal plants contain only low amounts of medicarpin.Moreover,the planting area for medicarpin-producing plants is limited;consequently,the current medicarpin supply cannot meet the high demands of medicinal markets.In this study,eight key genes involved in medicarpin biosynthesis were identified using comparative transcriptome and bioinformatic analyses.In vitro and in vivo enzymatic reaction confirmed the catalytic functions of candidate enzymes responsible for the biosynthesis of medicarpin and medicarpin intermediates.Further engineering of these genes in Saccharomyces cerevisiae achieved the heterologous biosynthesis of medicarpin using liquiritigenin as a substrate,with a final medicarpin yield of 0.82±0.18 mg/L.By increasing the gene copy numbers of vestitone reductase(VR)and pterocarpan synthase(PTS),the final medicarpin yield was increased to 2.05±0.72 mg/L.This study provides a solid foundation for the economic and sustainable production of medicarpin through a synthetic biology strategy.
基金supported by the National Natural Science Foundtion of China(No.81973194)CAMS Innovation Fund for Medical Sciences(CIFMS)(2019-12M-1-005).
文摘Main observation and conclusion Five novel pterocarpan derivatives(1—5)and three known pterocarpans were isolated from the roots of Sophora flavescens Ait.Their structures were elucidated based on extensive spectroscopic data(UV,IR,1D and 2D NMR,and HR-ESI-MS).Notably,compounds 1—5 were the first reported natural products,which possess unique pterocarpan-glucose-long chain aliphatic acid structures.The kinds of aliphatic acids were determined by comparison with authentic aliphatic acids using HPLC-ELSD analysis after acid hydrolysis.The absolute configurations of 1—5 were confirmed by analysis of their ECD spectra.All compounds were evaluated against lipopolysaccharide(LPS)-induced TNF-αproduction in RAW264.7 cells,and compound 1 showed significant inhibitory activity with inhibition rate of 86.22%,IC_(50)=0.79μmol/L(dexamethasone as positive control,inhibition rate of 71.38%).
