[Chemical constituents from ethyl acetate-soluble extraction of Valeriana jatamansi].

Application of a combination of various chromatographic techniques including column chromatography over silica gel, Sephadex LH-20, macroporous adsorbent resin, and reversed-phase HPLC, led to the isolation of 173 compounds including irdidoids, monoterpenes, sesquiterpenes, triterpenes, lignans, flavonoids, and simple aromatic derivatives from the ethyl acetate-soluble fraction of the whole plants of Valeriana jatamansi(Valerianaceae), and their structures were elucidated by spectroscopic methods including 1D, 2D NMR UV, IR, and MS techniques. Among them, 77 compounds were new. In previous reports, we have described the isolation, structure elucidation, and bioactivities of 68 new and 25 known compounds. As a consequence, we herein reported the isolation and structure elucidation of the remaining 9 new and 71 known compounds, the structure revision of valeriotriate A(8a), as well as cytotoxicity of some compounds.

[Pharmacokinetics of five ginsenosides of Shexiang Baoxin pill in plasma of myocardial infarction rats].

Shexiang Baoxin pill(SBP) is widely used for treating coronary heart disease in clinic, with ginsenosides as its main effective component. This study was designed to investigate and compare the pharmacokinetic characteristics of five ginsenosides of five compounds after multiple oral administrations, ginseng extract(GE) and SBP in myocardial infarction rats. After intragastric administration to myocardial infarction rats, the plasma samples were analyzed by liquid chromatography tandem triple-quad mass spectrometry. The results showed that Cmax of five compounds in all groups were less than 200 µg•L⁻¹. Tmax of corresponding analytes between groups revealed that ginsenosides in both SBP and GE were absorbed faster than each of the five compounds, indicating that GE and compounds contain components promoting absorption of ginsenosides. The oral administration doses of ginsenosides in SBP were the least in all groups, but with the greatest AUC0-tand AUCINF, which indicated that ginsenosides in SBP had the best absorption in all groups after oral administration to myocardial infarction rats. This study also demonstrated that compound is the best form of traditional Chinese medicine.

Three decomposition products of valepotriates from Valeriana jatamansi and their cytotoxic activity.

Three new decomposition products of valepotriates, valtrals A-C (1-3), and two known products, baldrinal and homobaldrinal, are formed during the isolation procedure of the ethanol extract of the whole plants of Valeriana jatamansi. Their structures were determined by spectroscopic methods including IR, MS, 1D, and 2D NMR experiments. Compounds 1-3 showed selective cytotoxicity against metastatic prostate cancer (PC-3M) and colon cancer (HCT-8) cell lines.

Efficient total synthesis and biological activities of 6-deoxyisojacareubin.

6-Deoxyisojacareubin was directly synthesized in a six-step route with an overall yield of about 20%. In this route, the excellent site selectivity of this Claisen rearrangement-cyclization reaction cascade was achieved by inserting a bulky p-tosyl group into the free 1-OH, and in the last step, some efficient demethylation methods were explored. Furthermore, all synthesized intermediates including 6-deoxyisojacareubin were evaluated for their inhibitory activity against the QGY-7703 cell line. Of these, compound 1 and 6-deoxyisojacareubin showed moderate activities with IC50 values of 39.61 and 9.65 µM, respectively, when compared to the positive control 5-fluorouracil with an IC50 value of 11.24 µM. Further investigation using non-radioactive detection of protein kinase C (PKC) suggested that these two compounds possessed potency in the inhibition of PKC.

Direct Identification and Quantitation of Protein Peptide Powders Based on Multi-Molecular Infrared Spectroscopy and Multivariate Data Fusion.

Given that protein peptide powders (PPPs) from different biological sources were inherited with diverse healthcare functions, which aroused adulteration of PPPs. A high-throughput and rapid methodology, united multi-molecular infrared (MM-IR) spectroscopy with data fusion, could determine the types and component content of PPPs from seven sources as examples. The chemical fingerprints of PPPs were thoroughly interpreted by tri-step infrared (IR) spectroscopy, and the defined spectral fingerprint region of protein peptide, total sugar, and fat was 3600-950 cm-1, which constituted MIR finger-print region. Moreover, the mid-level data fusion model was of great applicability in qualitative analysis, in which the F1-score reached 1 and the total accuracy was 100%, and a robust quantitative model was established with excellent predictive capacity (Rp: 0.9935, RMSEP: 1.288, and RPD: 7.97). MM-IR coordinated data fusion strategies to achieve high-throughput, multi-dimensional analysis of PPPs with better accuracy and robustness which meant a significant potential for the comprehensive analysis of other powders in food as well.

Reprogramming of cell junction modules during stepwise epithelial to mesenchymal transition and accumulation of malignant features in vitro in a prostate cell model.

Epithelial to mesenchymal transition (EMT) is pivotal in tumor metastasis. Our previous work reported an EMT model based on primary prostate epithelial cells (EP156T) which gave rise to cells with mesenchymal phenotype (EPT1) without malignant transformation. To promote prostate cell transformation, cells were maintained in saturation density cultures to select for cells overriding quiescence. Foci formed repeatedly following around 8 weeks in confluent EPT1 monolayers. Only later passage EPT1, but not EP156T cells of any passage, could form foci. Cells isolated from the foci were named EPT2 and formed robust colonies in soft agar, a malignant feature present neither in EP156T nor in EPT1 cells. EPT2 cells showed additional malignant traits in vitro, including higher ability to proliferate following confluence, higher resistance to apoptosis and lower dependence on exogenous growth factors than EP156T and EPT1 cells. Microarray profiling identified gene sets, many of which belong to cell junction modules, that changed expression from EP156T to EPT1 cells and continued to change from EPT1 to EPT2 cells. Our findings provide a novel stepwise cell culture model in which EMT emerges independently of transformation and is associated with subsequent accumulation of malignant features in prostate cells. Reprogramming of cell junction modules is involved in both steps.

New sesquiterpenoids from Ainsliaea macrocephala and their nitric oxide inhibitory activity.

Investigation of the ethanol extract of the whole plant of Ainsliaea macrocephala led to the isolation of five new sesquiterpenoids, namely ainsliadimer C (1), ainsliadimer D (2), ainsliaolide B (3), ainsliatone B (4), and ainsliaolide C (5), together with seventeen known sesquiterpenes and sesquiterpene glycosides (6- 22). Their structures were elucidated by spectroscopic methods. The relative stereochemistry of ainsliadimers C (1) and D (2) were further confirmed by single crystal X-ray diffraction analysis. Total extract of A. macrocephala and compounds 1- 22 were tested for inhibitory activity against the production of nitric oxide in RAW 264.7 cells stimulated by LPS, as well as for cytotoxicity against RAW 264.7 macrophages. Of all samples tested, purified compounds 4, 7, and 12 strongly inhibited the production of nitric oxide with IC50 values of 8.78, 2.50, and 7.11 µM, and simultaneously showed low cytotoxicity against RAW 264.7 macrophages.

Screening and analysis of the multiple absorbed bioactive components and metabolites in rat plasma after oral administration of Jitai tablets by high-performance liquid chromatography/diode-array detection coupled with electrospray ionization tandem mass spectrometry.

Based on the serum pharmacochemistry technique and high-performance liquid chromatography/diode-array detection (HPLC/DAD) coupled with electrospray tandem mass spectrometry (HPLC/ESI-MS/MS), a method for screening and analysis of the multiple absorbed bioactive components and metabolites of Jitai tablets (JTT) in orally dosed rat plasma was developed. Plasma was treated by methanol precipitation prior to liquid chromatography, and the separation was carried out on a Symmetry C(18) column, with a linear gradient (0.1% formic acid/water/acetonitrile). Mass spectra were acquired in negative and positive ion modes, respectively. As a result, 26 bioactive components originated from JTT and 5 metabolites were tentatively identified in orally dosed rat plasma by comparing their retention times and MS spectra with those of authentic standards and literature data. It is concluded that an effective and reliable analytical method was set up for screening the bioactive components of Chinese herbal medicine, which provided a meaningful basis for further pharmacology and active mechanism research of JTT.

Revision of the Structures of 1,5-Dihydroxy-3,8-epoxyvalechlorine, Volvaltrate B, and Valeriotetrate C from Valeriana jatamansi and V. officinalis.

The structures of 1,5-dihydroxy-3,8-epoxyvalechlorine (1a) and volvaltrate B (6a), two new chlorinated iridoids isolated from Valeriana jatamansi and V. officinalis, respectively, were originally assigned on the basis of spectroscopic methods. Reinvestigation using X-ray analysis and chemical transformation revealed that the original assignment of H-7 in 1a and OH-8 in 6a should be inverted and that the structures should be revised to 1 and 6, respectively. Correspondingly, the structure of valeriotetrate C (7a) should be revised to 7. Volvaltrate B (6) showed cytotoxic activity against the lung adenocarcinoma (A549), metastatic prostate cancer (PC-3M), colon cancer (HCT-8), and hepatoma (Bel7402) cell lines, with IC50 values of 8.5, 2.0, 3.2, and 6.1 µM, respectively.

Iridoids and lignans from Valeriana jatamansi.

Thirteen new iridoids including seven iridolactones, jatamanins A-M (1-13), and a new lignan, (+)-9'-isovaleroxylariciresinol (14), together with seven known iridoids and 13 lignans were obtained from whole plants of Valeriana jatamansi. Structures of the new compounds were determined by spectroscopic and crystallographic methods, and the absolute configuration of compound 1 was assigned by application of the modified Mosher method. Jatamanins H (8) and I (9) are iridolactones with an unusual C-8-C-11 oxygen bridge, forming a cage-like structure. (+)-9'-Isovaleroxylariciresinol (14) showed significant in vitro cytotoxicity against PC-3M and HCT-8 cell lines, with IC(50) values of 8.1 and 5.3 microM, respectively.

Antitumor and antiplatelet activity of alkaloids from veratrum dahuricum.

Ten steroidal alkaloids - cyclopamine, veratramine, jervine, 3, 15-diangyloylgermine, 3-angyloylzygadenine, 3-veratroyl zygadenine, 15-veratroylgermine, germine, veratrosine and pseudojervine - from Veratrum dahuricum, together with the ethanol extract and total alkaloids, were evaluated for their antitumor and antiplatelet activities. Cyclopamine, veratramine and germine significantly inhibited the hedgehog pathway in NIH/3T3 cells. Cyclopamine exerted a potent inhibitory effect against the growth of PANC-1 tumors in mice, with inhibition rates of 40.64%, 44.37%, 46.77% at doses of 5.0, 15.0 and 50.0 mg kg-1, respectively. Veratroylgermine was found to produce the strongest inhibition against the platelet aggregation induced by arachidonic acid, with inhibition rate of 92.0% at 100 microM.

Acylated iridoids with cytotoxicity from Valeriana jatamansi.

Thirteen new acylated iridoids, jatamanvaltrates A-M (1-13), together with nine known valepotriates (14-22), were isolated from the whole plants of Valeriana jatamansi (syn. Valeriana wallichii). The structures of these new compounds were assigned by detailed interpretation of spectroscopic data. Jatamanvaltrates D (4) and H (9) are the first examples of naturally occurring valepotriates containing an o-hydroxybenzoyloxy moiety at C-10. All isolated compounds were tested for their cytotoxicity against lung adenocarcinoma (A549), metastatic prostate cancer (PC-3M), colon cancer (HCT-8), and hepatoma (Bel7402) cell lines.

Constituents of the moss Polytrichum commune.

Two unusual flavonones coupled with styryl units, communins A (1) and B (2), and a new benzonaphthoxanthenone, ohioensin H (3), together with 11 known compounds, were isolated from the moss Polytrichum commune. The structures of 1-3 were assigned by spectroscopic data interpretation. The new compounds were evaluated for cytotoxicity against a small panel of cancer cell lines.

cis-Clerodane diterpenoids from the liverwort Gottschelia schizopleura and their cytotoxic activity.

Two new and three previously known CIS-clerodane diterpenoids were isolated from the wild liverwort Gottschelia schizopleura (Jungermanniales, Jungermanniaceae). Their structures were established on the basis of spectroscopic analysis, especially 1D and 2D NMR data. The cytotoxic activities of compounds 1- 5 were evaluated against liver hepatoblastoma (HEP-G2), lung carcinoma (A549), breast ductal carcinoma (MDA-MB-435), and colon adenocarcinoma (LOVO) cell lines. Compound 1 showed moderate inhibition against MDA-MB-435 and LOVO cells.

Chemical constituents of Crinum asiaticum L. var. sinicum Baker and their cytotoxic activities.

A phytochemical investigation of the bulbs of Crinum asiaticum L. var. sinicum Baker resulted in the isolation of two new alkaloids, asiaticumines A and B (1 and 2, resp.), together with 21 known compounds, including nine alkaloids, four amides, five phenolic compounds, and three flavonoids. All 23 compounds were isolated for the first time from Crinum asiaticum L. var. sinicum Baker. Their structures were elucidated by spectroscopic methods. In addition, ten alkaloids, 1-10, were evaluated for their cytotoxic activities against human tumor cell lines A549, LOVO, HL-60, and 6T-CEM. Compounds 3, 4, and 7-10 selectively showed remarkable inhibition against one or more of the tested cell lines.

Antitumor activity of extracts and compounds from the rhizomes of Veratrum dahuricum.

The antitumor activity of six extracts (ethanol extract, petroleum ether fraction, CHCl(3) fraction, ethyl acetate fraction, n-butanol fraction and total alkaloids) from the rhizomes of Veratrum dahuricum, and six compounds (veratramine (1), jervine (2), germine (3), veramitaline (4), veratrosine (5) and cyclopamine (6)) from the ethanol extract were investigated in vitro. The 12 samples exhibited cytotoxic activity against human tumor cell lines A549, PANC-1, SW1990 and NCI-H249. Among these samples, CHCl(3) fraction, the total alkaloids, compounds 1 and 6 showed higher inhibitory activity, compound 3 selectively exhibited significant cytotoxicity to SW1990 and NCI-H249.

Alkaloids from Corydalis saxicola and their anti-hepatitis B virus activity.

Eight protoberberine-type alkaloids and two indole alkaloids were isolated from the MeOH extracts of the herb Corydalis saxicola Bunting (Papaveraceae). Their structures were identified as dehydrocavidine (1), dehydroapocavidine (2), dehydroisoapocavidine (3), berberine (4), dehydroisocorypalmine (5), coptisine (6), tetradehydroscoulerine (7), berbinium (8), 1-formyl-5-methoxy-6-methylindoline (9), and 1-formyl-2-hydroxy-5-methoxy-6-methylindoline (10). Compounds 3, 9, and 10 are new alkaloids. All compounds were tested for anti-HBV activity against the 2.2.15 cell line in vitro. Dehydrocavidine (1), dehydroapocavidine (2), and dehydroisoapocavidine (3) exhibited inhibitory activity against HBsAg and HBeAg, but no cytotoxicity against the 2.2.15 cell line.

Quantitative determination of the anticancer agent tubeimoside I in rat plasma by liquid chromatography coupled with mass spectrometry.

Tubeimoside I is an important component isolated from Bolbostemma paniculatum. Tubeimoside I has been demonstrated to possess many pharmacological activities, including anti-inflammatory, antitumor, and antitumor-promoting effects. The purpose of the present study was to examine in vivo pharmacokinetics and bioavailability of tubeimoside I in rats by using a liquid chromatography coupled with mass spectrometry quantitative detection method (LC/MS). The plasma samples were deproteinated, evaporated and reconstituted in 100 microl methanol prior to analysis. The separation was performed by Waters Symmetry C18 reversed-phase column (3.5 microm, 150 mm x 2.1mm, Waters Inc., USA) and a SB-C18 guard column (5 microm, 20 mm x 4.0mm). The mobile phase was a mixture of acetonitrile and water containing 5 microM NaAc (60:40, v/v). The method was validated within the concentration range 20-5000 ng/ml, and the calibration curves were linear with correlation coefficients >0.999. The lowest limit of quantitation (LLOQ) for tubeimoside I was 20 ng/ml in 0.1 ml rat plasma. The intra-assay accuracy and precision ranged from 92.4 to 104.9% and from 5.8 to 10.5%, respectively, while inter-assay accuracy and precision ranged from 94.2 to 95.0% and from 5.1 to 8.8%, respectively. The method was further applied to assess pharmacokinetics and oral bioavailability of tubeimoside I after intravenous and oral administration to rats. The oral bioavailability of tubeimoside I is only 0.23%, which indicates that tubeimoside I has poor absorption or undergoes acid-induced degradation. Practical utility of this new LC/MS method was confirmed in pilot pharmacokinetic studies in rats following both intravenous and oral administration.

A new bisxanthone from Hypericum japonicum.

A new bisxanthone, named jacarelhyperols D (1), was isolated from the whole plant of Hypericum japonicum. Its chemical structure was elucidated as 6-[1',5',6'-trihydroxy-2''-(alpha-hydroxy-alpha-methyl)ethyl-3'',4''-dihydrofuran (2'',3'',3',4') xanthone-3''-oxyl]-1,3,5-trihydroxy-xanthone on the basis of spectroscopic analysis.