Anti-listerial compounds from Asari Radix

Asari Radix, the roots of Asarum heterotropoides F. Maekawa var. manshuricum F. Maekawa or A. sieboldii F. Maekawa, has traditionally been used for the treatment of various infectious diseases. Since its MeOH extract inhibited the growth of Listeria monocytogenes in a preliminary test, the aim of this study was to isolate and identify the anti-listerial compounds from the plant. Activity-guided fractionation led to the isolation of seven compounds 1–7 from the MeOH extract, and their chemical structures were identified by comparison of the spectroscopic data with those in the literature. Compounds 1–7 exhibited inhibitory activity against all five tested strains of L. monocytogenes with diameter of inhibition zones ranging from 7 to 11 mm in the agar disc diffusion method. Compounds 1–3 and 7 demonstrated potent antimicrobial effects on the L. monocytogenes strains, with MICs between 62.5 and 125 μg/mL. This is the first report that AR possesses inhibitory activity against L. monocytogenes.     

Analysis of aristolochic acid in nine sources of Xixin, a traditional Chinese medicine, by liquid chromatography/atmospheric pressure chemical ionization/tandem mass spectrometry

Aristolochic acid I (AA-I), which is a known nephrotoxin, is found in a commonly used Chinese medicine, Xixin, that originates from nine Asarum species (Aristolochiaceae) found in China. A method has been developed using reversed-phase liquid chromatography coupled with atmospheric pressure chemical ionization (APCI) tandem mass spectrometry under the positive ion detection mode [LC/(+)APCI/MS/MS] to determine the amount of AA-I in Xixin. The limit of detection of AA-I, estimated by monitoring with LC/MS/MS, was at the low microg/l level. By applying this method to methanol extracts of nine Asarum species, the concentrations of AA-I were found to range from 3.3 ng/mg (Asarum sieboldii) to 3376.9 ng/mg (Asarum crispulatum).     

UPLC-UV测定细辛地上和地下部位中的马兜铃酸A

马兜铃酸因被证实具有肾毒性而引起广泛的重视[1,2],尤其近年来美国FDA有关法令中,已明确提出禁止使用含有马兜铃酸的药物,因此马兜铃科植物的安全性受到质疑.     

Toxicities of aristolochic acid I and aristololactam I in cultured renal epithelial cells

Aristolochic acid nephropathy, a progressive tubulointerstitial renal disease, is primarily caused by aristolochic acid I (AA-I) intoxication. Aristololactam I (AL-I), the main metabolite of AA-I, may also participate in the processes that lead to renal damage. To investigate the role and mechanism of the AL-I-mediated cytotoxicity, we determined and compared the cytotoxic effects of AA-I and AL-I on cells of the human proximal tubular epithelial (HK-2) cell line. To this end, we treated HK-2 cells with AA-I and AL-I and assessed the cytotoxicity of these agents by using the 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay, flow cytometry, and an assay to determine the activity of caspase 3. The proliferation of HK-2 cells was inhibited in a concentration- and time-dependent manner. Cell-cycle analysis revealed that the cells were arrested in the S-phase. Apoptosis was evidenced by the results of the annexin V/propidium iodide (PI) assay and the occurrence of a sub-G1 peak. In addition, AA-I and AL-I increased caspase 3-like activity in a concentration-dependent manner. These results also suggested that the cytotoxic potency of AL-I is higher than that of AA-I and that the cytotoxic effects of these molecules are mediated through the induction of apoptosis in a caspase 3-dependent pathway.     

HPLC analysis of coumarins in Turkish <Emphasis Type="Italic">Seseli</Emphasis> species (Umbelliferae)

The n-hexane extracts of aerial and underground parts of three Seseli species, S. gummiferum subsp. corymbosum, S. resinosum, and S. hartvigii growing in Turkey were investigated concerning the presence of coumarins by normal-phase HPLC (high-performance liquid chromatography). In this respect, two coumarins, osthol (1) and corymbocoumarin (2) [(−)-(3′S, 4′S)-3′-acetoxy-4′-isovaleryloxy-3′, 4′-dihydroseselin] isolated from aerial parts of S. gummiferum subsp. corymbosum, were used as standard samples. The amount of these compounds were discussed and evaluated in this study.     

Simple and rapid analysis of aristolochic acid contained in crude drugs and Kampo formulations with solid-phase extraction and HPLC photodiode-array detection

Simple and rapid analysis of aristolochic acid (AA) in crude drugs and Kampo extracts using a solid-phase extraction method and HPLC-PDA analysis was investigated. Extraction of AA from samples was accomplished by adding methanol containing 1% ammonia. The addition of ammonia ionized the AA of acidic substances so that they adhered to an acrylamide copolymer of a strong anion exchange resin (Sep-Pak QMA) coupled to diol silica easily. Furthermore, a mixture of acetonitrile–water–phosphoric acid (75:25:2, v/v) was effective in isolating AA from its carrier. Since almost all interfering peaks originating from contaminants in crude drugs and Kampo extract formulations could be removed, a satisfactory HPLC chromatogram of AA was obtained. A good result was also obtained when Aristolochiaceae and crude drugs containing AA were tested. Particularly in the case of the medicinal parts of Asarum, several interfering peaks and a ghost peak detected near the AA peak were eliminated. The AA contents of two Kampo extract formulations, tokishigyakukagoshuyushokyoto and ryutanshakanto, were calculated by HPLC analysis. The AA content (the sum of AA-I and AA-II) was 1.25–6.13 mg per daily dose. From an additional recovery experiment for Kampo formulations, high recovery rates of AA were obtained. Neither LC/MS nor special instrumentation was necessary. Our results suggest that this simple, quick, and sensitive analytical method to detect AA in crude drugs and Kampo extract formulations would be valuable in safety inspections of AA in crude drugs and their products.     

辽细辛地上部分化学成分的分离与鉴定

目的对辽细辛(Asarum heterotropidesFr Schmidt.var.mandshuricum(Maxim.)Kitag.)地上部分的化学成分进行研究。方法采用硅胶柱色谱等方法进行分离纯化,根据理化性质和NMR波谱数据进行结构鉴定。结果分离得到6个化合物,分别鉴定为(10bS)8,9-dihydroxy-1,5,6,10b-tetrahydro-2H-pyrrolo[2,1-a]isoquinolin-3-one(1)、尼古丁酸(nicotinic acid,2)、5-羟甲基糠酸(5-hy-droxymethyl-furoic acid,3)、琥珀酸(succinic acid,4)、山奈酚(kaempferol,5)、山奈酚3-O--βD-吡喃葡萄糖苷(kaempferol-3-O--βD-glucopyranoside,6)。结论化合物1~3为首次从细辛属植物中分离得到,化合物5为首次从辽细辛中分离得到。     

A new furofuran lignan from <Emphasis Type="Italic">Isodon japonicus</Emphasis>

A new sesamin type furofuran lignan, (−)-sesamin-2,2′-diol (1), along with two known flavonoids (2 and 3) and three phenolic compounds (4–6) were isolated from the aerial parts of Isodon japonicus. The structures of these compounds were determined by analysis of spectroscopic data (1D-, 2D-NMR, HRMS and CD) and by comparison of the data with those of related metabolites.     

Chemical characteristics for different parts of Panax notoginseng using pressurized liquid extraction and HPLC-ELSD

The chemical characteristics for different parts of Panax notoginseng, including root, fibre root, rhizome, stem, leaf, flower and seed, were determined using high performance liquid chromatography-evaporative light scattering detection (HPLC-ELSD) and pressurized liquid extraction (PLE). Eight major saponins, namely notoginsenoside R1, ginsenosides Rg1, Re, Rb1, Rc, Rb2, Rb3 and Rd were also quantitatively compared among the different parts of P. notoginseng. The chromatograms showed that there was significant difference between underground (root, fibre root, rhizome) and aerial (leaf and flower) parts from P. notoginseng, though the similarities of entire chromatographic patterns among tested samples from underground (0.965 ± 0.029, n = 12) and aerial parts (0.987 ± 0.014, n = 5) were similar, respectively. Especially, no saponin was detected in the seed of P. notoginseng. Hierarchical clustering analysis based on eight investigated saponins or the ratios of contents for ginsenoside Rg1/Rb1 and ginsenoside Rb3/Rb1 showed that the samples from different parts of P. notoginseng were divided into three main clusters. One cluster was underground parts, which contained rich protopanaxatriol and protopanaxadiol types saponins. The leaf and flower were in the same cluster, which contained protopanaxadiol type saponins only. Especially, ginsenoside Rc, Rb2 and Rb3, rare in the underground parts, were rich in aerial parts of P. notoginseng. The stem of P. notoginseng was another cluster. Based on the cluster analysis, the chemical characteristics for different parts of P. notoginseng were revealed. They are composite cluster (underground parts), protopanaxadiol cluster (aerial parts) and interim (stem) cluster, which was the one between the two typical clusters, respectively. The result shows that chemical characteristics of underground parts and aerial parts from P. notoginseng are obviously different, which is helpful for pharmacological evaluation and quality control of P. notoginseng.     

Aromatic compounds and triterpenoidal saponins fromClematis koreana var.umbrosa

From the methanolic extract of aerial parts ofClematis koreana var.umbrosa, one new triterpenoidal saponin, 3-O-β-D-xylopyranosyl(1–3)-α-L-arabinopyranosyl oleanolic acid 28-O-α-L-rhamnopyranosyl(1–4)-β-D-glucopyranosyl(1–6)-β-D-glucopyranosyl ester, along with five known aromatic compounds and two known triterpenoidal saponins were isolated.     

高效液相色谱法检测北细辛药材不同部位的马兜铃酸

目的检测北细辛药材不同部位的马兜铃酸含量。方法采用高效液相色谱（HPLC）法,色谱柱为C18柱（150mm×4.6mm,5μm）,磷酸二氢钠-乙腈（60：40）为流动相,检测波长为400nm。结果北细辛药材地上部分、地下部分及全草中均未检出马兜铃酸A。结论该法简便、快速、准确,可用于北细辛药材中马兜铃酸的检测。     

Biologically active <Emphasis Type="Italic">C</Emphasis>-alkylated flavonoids from <Emphasis Type="Italic">Dodonaea viscosa</Emphasis>

A new C-alkylated flavonoid (5,7-dihydroxy-3′-(4″-acetoxy-3″-methylbutyl)-3,6,4′-trimethoxyflavone (1), along with two known C-alkylated flavonoids (5,7-dihydroxy-3′-(3-hydroxymethylbutyl)-3,6,4′-trimethoxyflavone (2), 5,7,4′-trihydroxy-3′-(3-hyroxymethylbutyl)-3,6-dimethoxyflavone (3) and two new source C-alkylated flavonoids (5,7-dihydroxy-3′-(2-hydroxy-3-methyl-3-butenyl)-3,6,4′-trimethoxyflavone (4), 5,7,4′-trihydroxy-3,6-dimethoxy-3′-isoprenyl-flavone (5) were isolated from the aerial parts of Dodonaea viscosa. The structures of all compounds were established on the basis of 1D and 2D NMR spectroscopy and mass spectrometry. The isolated compounds were evaluated for their inhibitory effect on urease and α-chymotrypsin enzyme. All the compounds (1–5) exhibited mild inhibition against urease but remained recessive in case of α-chymotrypsin.     

Pharmacological and histological effects of <Emphasis Type="Italic">Centaurea bruguierana</Emphasis> ssp. <Emphasis Type="Italic">belangerana</Emphasis> on indomethacin-induced peptic ulcer in rats

The species Centaurea bruguierana (DC.) Hand.-Mazz. ssp. belangerana (DC.) Bornm. (CBB) (Asteraceae), known as “Baad-Avard” in Borazjan, Bushehr Province, southern Iran, is used in folk medicine as a hypoglycemic herb in diabetes and as a remedy for peptic ulcer disorders. Total 80% EtOH extract and petroleum ether, CHCl₃, EtOAc, n-BuOH, and remaining fractions obtained by solvent–solvent fractionation of dried aerial flowering parts of the plant were investigated for anti-ulcer activity against indomethacin-induced ulcerogenesis in rats. Anti-ulcer activity was evaluated by measuring the ulcer index (UI) and ulcer inhibition. The UI was significantly reduced in all treated animals. A dramatic decrease in the UI was observed following the administration of total extract (100 mg/kg, p < 0.001) and CHCl₃ fraction (42 mg/kg, ***p < 0.001) in comparison with the control group. The percentage ulcer inhibition with total extract at a dose of 100 mg/kg (97.66%) and CHCl₃ fraction at a dose of 42 mg/kg (96.96%) was found to be higher (p < 0.001) than the reference group (cimetidine 100 mg/kg) (87.08%). The pharmacological and histological results of the present study proved that the aerial flowering parts of CBB possess preventive activity against peptic ulcer, supporting the traditional assertion in southern Iranian folk medicine.     

Aporphine alkaloids from <Emphasis Type="Italic">Clematis parviloba</Emphasis> and their antifungal activity

A new aporphine alkaloid, β-magnoflorine (1), together with a known aporphine alkaloid, α-magnoflorine (2), were isolated from the aerial parts of Clematis parviloba. Their structures were elucidated on the basis of comprehensive spectroscopic techniques. In addition, both compounds showed potent antifungal activities against Penicillium avellaneum UC-4376.     

辽细辛地下部分的化学成分(Ⅱ)

目的对辽细辛地下部分进行化学成分研究。方法采用反复硅胶柱色谱法分离纯化,通过理化性质和谱学分析鉴定化合物结构。结果分离得到6个化合物,分别鉴定为马兜铃酰胺Ⅰ(aristololactamⅠ,1)、7-甲氧基马兜铃酰胺(7-methoxy-aristololactam,2)、马兜铃酸Ⅳa(aristolochicacidⅣa,3)、对羟基苯甲酸(p-hydroxybenzoic acid,4)、5,7-二-O-β-D-吡喃葡萄糖基柚皮素(5,7-di-O-β-D-glucpyranosyl-narigenin,5)、2,6-二甲氧基-4-甲基苯基-1-O-β-D-吡喃葡萄糖苷(2,6-dime-thoxy-4-methylphenyl-1-O-β-D-glucopyranoside,6)。结论化合物4、6为细辛属内首次分离得到,化合物2、5为从该植物中首次分离得到。     

A new abietane diterpenoid from Isodon inflexus

A new ent-abietane diterpenoid, 3α,6β-dihydroxy-7,17-dioxo-ent-abieta-15(16)-ene (1), and three known ent-kaurane diterpenids, kamebacetal A (2), kamebakaurin (3), and excisanin A (4), and a known triterpenoid, ursolic acid (5), were isolated from the aerial parts of Isodon inflexus. Their chemical structures were determined by extensive analysis of spectroscopic data including 1D-and 2D-NMR experiments. All isolates (1–5) were evaluated for their potential to inhibit LPS-induced nitric oxide production in RAW264.7 cells. Of these, compounds 1–4 inhibited the production of NO with IC₅₀ values ranging from 1.0 to 26.5 μM.     

Megastigmane glucoside from <Emphasis Type="Italic">Asystasia gangetica</Emphasis> (L.) T. Anderson

A 5,11-epoxymegastigmane glucoside (asysgangoside) was isolated from the aerial parts of Asystasia gangetica together with the known compounds, salidroside, benzyl β-d-glucopyranoside, (6S,9R)-roseoside, ajugol, apigenin 7-O-β-d-glucopyranoside, apigenin 7-O-neohesperidoside, and apigenin 7-O-β-d-glucopyranosyl (1→6)-β-d-glucopyranoside. The structure elucidations were based on spectroscopic evidence.     

New anthracene glycosides from <Emphasis Type="Italic">Rhodomyrtus tomentosa</Emphasis> stimulate osteoblastic differentiation of MC3T3-E1 cells

Two new anthracene glycosides (1, 2) were isolated from aerial parts of Rhodomyrtus tomentosa, along with three known compounds (3–5). The structures of two new compounds were established to be 4,8,9,10-tetrahydroxy-2,3,7-trimethoxyanthracene-6-O-β-D-glucopyranoside (1) and 2,4,7,8,9,10-hexahydroxy-3-methoxyanthracene-6-O-α-L-rhamnopyranoside (2) based on spectroscopic and chemical methods. Among them, compound 1, 2, and 5 significantly (P<0.05) increased the alkaline phosphatase activity, collagen synthesis, and mineralization of the nodules of MC3T3-E1 osteoblastic cells compared to those of the control, respectively.     

Technology for cinaroside production from the aerial part of <Emphasis Type="Italic">Ferula varia</Emphasis> and evaluation of its hypoazotemic activity

The optimal regime of cinaroside extraction from the aerial part of Ferula varia has been found. Aprocessing scheme for obtaining cinaroside has been developed on the basis of these data. Two samples of cinaroside (obtained by the laboratory and semi-industrial methods) are comparable to the well-known medicinal preparation lespenephryl with respect to hypoazotemic action. This fact indicates that Ferula varia is a valuable source of cinaroside. A technology has been developed for producing a new medicinal form of cinaroside in 0.05 g tablets. Translated from Khimiko-Farmatsevticheskii Zhurnal, Vol. 43, No. 3, pp. 38 – 40, March, 2009.