Rubochingosides A - J, labdane-type diterpene glycosides from leaves of Rubus chingii.

Ten previously undescribed labdane-type triterpenoids, rubochingosides A - J (1-10), were isolated from the leaves of Rubus chingii Hu. Their structures were elucidated by spectroscopic and chemical methods. The cytotoxicity of all isolated compounds was tested against five human tumor cell lines (Bel-7402, Caski, BGC-823, A2780, and HCT-116). Among them, compounds 4 and 8 showed cytotoxic activities against Bel-7402 and A2780 cells with IC50 values of 10.43 ± 0.51 and 29.03 ± 2.94 µM, respectively; Compound 8 exhibited cytotoxic activities against Bel-7402 and HCT-116 cells with IC50 values of 17.78 ± 1.54 and 26.23 ± 6.14 µM, respectively.

Ultra-high-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry coupled with three-step data post-processing techniques for comprehensive profiling of the multiple components in Fufang Xianzhuli Ye.

INTRODUCTION: Fufang Xianzhuli (FXZL) Ye, a classical formula of traditional Chinese medicine, is composed of Succus Bambusae, Houttuyniae herba, Pinelliae Rhizoma, Zingiberis Rhizoma Recens, Eriobotryae Folium, Platycodonis Radix, and peppermint oil. For many years, FXZL has been primarily utilised in China to treat cough and phlegm. The chemical composition of FXZL has not been reported, which seriously affects the safety of the clinical application. OBJECTIVE: To establish a systematic method for rapidly classifying and recognising the chemical constituents in the FXZL for the safety of the clinical application. METHODS: An ultra-high performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry coupled with a three-step data post-processing strategy was developed to screen the chemical constituents of FXZL. RESULTS: In this experiment, the diagnostic ions in FXZL were classified into six main compounds. A total of 106 compounds were unambiguously identified in FXZL based on their retention times, accurate masses, and tandem mass spectrometry data. These include 11 chlorogenic acids, three flavonoids, eight sesquiterpenoids, six organic acids, 65 triterpenoid saponins, and 13 other compounds. CONCLUSION: The chemical composition of FXZL was identified and summarised, providing useful information for quality control and a basis for further exploration of its active ingredients in vivo.

Identification and quantification of five impurities in cloperastine hydrochloride.

Cloperastine hydrochloride, a piperidine derivative, is a drug substance with a central antitussive effect and widely used in cough treatment; and its impurities have not been reported. Herein we isolated and identified five impurities (named as impurity A, B, C, D and E) in cloperastine hydrochloride bulk drug and developed a quantitative HPLC method. First, impurity A, B, C were enriched by ODS column chromatography and isolated by semi-preparative HPLC, at the same time, impurity D was purified by ODS column chromatography. Then, impurity E was enriched by strong acid degradation and purified by semi-preparative HPLC. At last, their structures were characterized by a variety of spectral data (MS, 1H NMR, 13C NMR, HSQC, HMBC and 1H-1H COSY). Impurity A was confirmed as 1-[2-(diphenylmethoxy)ethyl]piperidine, which having one less chloro-substituent compared with cloperastine. Impurity B was confirmed as 1-[2-[(2-chlorophenyl)(phenyl)methoxy]ethyl]piperidine, which was the isomer of cloperastine with 2-chloro-substituent. Impurity C was confirmed as 1-[2-[(3-chlorophenyl)(phenyl)methoxy]ethyl]piperidine, which was the isomer of cloperastine with 3-chloro-substituent. Impurity D was confirmed as (4-chlorophenyl)(phenyl)methanone, which was the raw material for the synthesis of cloperastine. Impurity E was confirmed as (4-chlorophenyl)(phenyl)methanol, which was an intermediate in the synthesis of cloperastine, and it was also a hydrolysate of cloperastine. Finally, the developed method was validated in terms of specificity, linearity, sensitivity, precision and accuracy.

Two new iridoid glycosides from Callicarpa nudiflora.

Two new iridoid glycosides, callicoside E (1) and callicoside F (2), were isolated from the leaves of Callicarpa nudiflora. Their structures were established by one- and two-dimensional NMR spectroscopy and mass spectrometry. In an in vitro bioassay, compounds 1 and 2 showed an pronounced hepatoprotective activity against d-galactosamine-induced toxicity in WB-F344 rat hepatic epithelial stem-like cells.[Formula: see text].

Two new norneolignans from Callicarpa kwangtungensis.

Two new norneolignans, (7S,8R)-3-methoxy-3',4,9-trihydroxy-4',7-epoxy-8,3'-neolignane-1'-carboxylic acid (1) and (7R,8R)-3-methoxyl-4,9-dihydroxy-3':7,4':8-diepoxyneolignan-1'-carboxylic acid methyl ester (2) were isolated from Callicarpa kwangtungensis, together with ten known compounds, genistin (3), daidzin (4), silybin A (5), isosilybin A (6), isosilybin B (7), p-hydroxybenzaldehyde (8), syringic acid (9), lanceolatin A (10), icariside C5 (11), and (3S,6E,10R)-10-ß-D-glucopyranosyloxy-3,11-dihydroxy-3,7,11-trimethyldodeca-1,6-diene (12). Compounds 1 and 2 were evaluated for their effects on the inhibition of nitric oxide (NO) production in lipopolysaccharide induced RAW264.7 cells. Compounds 1 and 2 exhibited inhibitory activity with IC50 values of 31.45 ± 0.38 and 40.72 ± 0.54 µM, respectively.

Two new noroleanane-type triterpenoid saponins from the stems of Stauntonia chinensis.

Two new noroleanane-type triterpenoid saponins, 3ß,20α,24-trihydroxy-29-norolean-12-en-28-oic acid 24-O-ß-L-fucopyranosyl-(1→2)-[ß-D-xylopyranosyl-(1→3)]-ß-D-glucopyranoside (1) and 3ß,20α,24-trihydroxy-29-norolean-12-en-28-oic acid 24-O-ß-D-glucopyranosyl-(1→2)-[α-L-arabinopyranosyl-(1→3)]-ß-D-glucopyranoside (2) were isolated from the stems of Stauntonia chinensis DC., together with three known compounds, brachyantheraoside B2 (3), eupteleasaponin Ⅷ (4) and fargoside B (5). Their structures were elucidated by spectroscopic and chemical methods. The cytotoxic activities of compounds 1 and 2 were evaluated against five human tumor cell lines (HCT-116, HepG2, BGC-823, NCI-H1650, and A2780). Compounds 1 and 2 showed moderate cytotoxic activities toward the tested cell lines with IC50 values ranging from 12.71 to 32.04 µM.

Inhibitory effects of luteolin­4'­O­ß­D­glucopyranoside on P2Y12 and thromboxane A2 receptor­mediated amplification of platelet activation in vitro.

Platelet activation and subsequent accumulation at sites of vascular injury are central to thrombus formation, which is considered to be a trigger of several cardiovascular diseases. Callicarpa nudiflora (C. nudiflora) Hook is a traditional Chinese medicinal herb for promoting blood circulation by removing blood stasis. In our previous study, several compounds extracted from this herb, including luteolin­4'­O­ß­D­glucopyranoside (LGP), were revealed to exert inhibitory effects on adenosine diphosphate (ADP)­induced platelet aggregation. The aim of present study was to confirm these antiplatelet effects and elucidate the potential mechanisms. Using a platelet­aggregation assay, it was revealed that LGP significantly inhibited platelet aggregation induced by ADP, U46619 and arachidonic acid. It was also found that LGP exhibited marked inhibitory effects on the activation of αIIbß3 integrin, the secretion of serotonin from granules, and the synthesis of thromboxane A2. In addition, the results showed that LGP suppressed Ras homolog family member A and phosphoinositide 3­kinase/Akt/glycogen synthase kinase 3ß signal transduction. Data from a radiolabeled ligand­binding assay indicated that LGP exhibited apparent competing effects on thromboxane receptor (TP) and P2Y12 receptors. In conclusion, the data presented here demonstrated that LGP, a natural compound from C. nudiflora Hook, inhibited the development of platelet aggregation and amplification of platelet activation. These inhibitory effects may be associated with its dual­receptor inhibition on P2Y12 and TP receptors.

Two new iridoid glycosides from the leaves of Callicarpa nudiflora.

Two new iridoid glycosides, callicoside C (1) and callicoside D (2), together with three known compounds (3-5), were isolated from the leaves of Callicarpa nudiflora. Their structures were established by 1D and 2D NMR spectroscopy and mass spectrometry. In an in vitro bioassay, compound 1 showed pronounced hepatoprotective activity against d-galactosamine-induced toxicity in WB-F344 rat hepatic epithelial stem-like cells.

Cytotoxic triterpenoid saponins from the defatted seeds of Camellia oleifera Abel.

Five new oleanane-type triterpenoid saponins, oleiferasaponins D1-D5 (1-5), were isolated from the defatted seeds of Camellia oleifera Abel. Their structures were elucidated by spectroscopic and chemical methods. The cytotoxic activities of compounds 1-5 were evaluated against five human tumor cell lines (HCT-116, HepG2, BGC-823, NCI-H1650, and A2780). Compounds 1-2 exhibited cytotoxic activity against five human cancer cell lines, with IC50 values ranging from 3.31 to 10.23 µM. Compounds 3-5 showed moderate cytotoxic activities toward the tested cell lines.

1, 6-di-O-caffeoyl-ß-D-glucopyranoside, a natural compound from Callicarpa nudiflora Hook impairs P2Y12 and thromboxane A2 receptor-mediated amplification of platelet activation and aggregation.

BACKGROUND: Platelet activation and subsequent accumulation at sites of vascular injury perform a central role in thrombus formation, which is believed to be the trigger of several cardiovascular diseases, such as atherosclerosis, myocardial infarction and strokes. In this sense, the search for agents that are capable of blocking platelets aggregation has important implications for these diseases. Callicarpa nudiflora (C. nudiflora) Hook is a traditional Chinese medicine herb for eliminating stasis to subdue swelling and hemostasis. Our previous study found several compounds extracted from this herb, including 1, 6-di-O-caffeoyl-ß-D-glucopyranoside (CGP), showed inhibitory effects on adenosine diphosphate (ADP) induced platelet aggregation. PURPOSE: The aim of current study is confirmation of the anti-platelet effects and elucidation of the probable mechanisms. METHODS: The experiments were performed on platelet rich plasma freshly isolated from SD rat. ADP, U46619 or arachidonic acid (AA) induced platelet aggregation assay were performed to evaluate the anti-platelet properties of CGP. Activated αIIbß3 integrin abundance, serotonin (5-HT) secretion, thromboxane A2 (TXA2) synthesis was determined to assess the effects of CGP on platelet activation. Furthermore, RhoA and PI3K/Akt/GSK3ß signal transduction were analyzed by Western Blotting assay. In addition, radiolabelled ligand binding assay was involved to evaluate the ability of CGP binding to thromboxane prostanoid (TP) and P2Y12 receptors. RESULTS: CGP inhibited platelet aggregation induced by ADP, U46619 and arachidonic acid (AA), significantly. Furthermore, it is also found that LGP exhibited obvious inhibitory effects on αIIbß3 integrin activation, serotonin (5-HT) secretion from granule and thromboxane A2 (TXA2) synthesis. Next, we found that CGP suppressed RhoA and PI3K/Akt/GSK3ß signal transduction. Data from radiolabelled ligand binding assay showed that CGP displayed apparent competing effects on TP and P2Y12 receptors. CONCLUSION: Collectively, the data presented here demonstrated that CGP, a natural compound from Callicarpa nudiflora Hook, inhibited the development of platelet aggregation and amplification of platelet activation. These inhibitory effects may be associated with its dual-receptor inhibition on P2Y12 and TP receptors.

Hepatoprotective iridoid glucosides from Callicarpa nudiflora.

Two new iridoid glucosides, callicoside A (1) and callicoside B (2), were isolated from the leaves of Callicarpa nudiflora. Their structures were elucidated by means of spectroscopic methods and chemical evidences. In an in vitro bioassay, compound 1 showed pronounced hepatoprotective activity against D-galactosamine-induced toxicity in WB-F344 rat hepatic epithelial stem-like cells.

Two new triterpenoid glycosides from the stems of Camellia oleifera Abel.

Two new oleanane-type triterpenoid glycosides, 3-O-ß-D-xylopyranosyl-(1→2)-α-L-arabinopyranosyl-(1→3)-[ß-D-glucuronopyranosyl-(1→2)]-ß-D-glucuronopyranosyl-22α-angeloyloxyolean-12-ene-15α,16α,28-triol(1) and 3-O-ß-D-xylopyranosyl-(1→2)-α-L-arabinopyranosyl-(1→3)-[ß-D-glucuronopyranosyl-(1→2)]-ß-D-glucuronopyranosyl-21ß-acetyl-22α-angeloyloxyolean-12-ene-16α,28-diol (2) were isolated from the stems of Camellia oleifera Abel. Their structures were elucidated by means of spectroscopic methods and chemical evidence. The cytotoxic activities of compounds 1-2 were evaluated against five human tumour cell lines (HCT-8, BGC-823, A5049, and A2780). Compounds 1-2 showed cytotoxic activity against five human cancer cell lines, with IC50 values ranging from 3.15 to 7.32 µM.

[Chemical Constituents from Stauntonia chinensis].

Objective: To study the chemical constituents of Stauntonia chinensis. Methods: The chemical constituents were isolated and purified by column chromatography on silica gel,ODS,Sephadex LH-20 and MPLC. Their structures were elucidated on the basis of physicochemical properties and special analysis. Results: Seven compounds were isolated from the leaves of Stauntonia chinensis,whose structures were elucidated as 3-O-ß-D-glucopyranosyl-( 1 →3)-[ß-D-xylopyranosyl-( 1 →2) ]-α-L-arabinopyranosyl-28-O-[α-L-rhamnopyranosyl-( 1 →4)-ß-D-glucopyranosyl-( 1 →6)-ß-D-glucopyranosyl]-3ß-hydroxy-30-norolean-12,20( 29)-dien-28-oic acid( 1),3-[( O-ß-D-glucopyranosyl-( 1→3)-[α-L-rhamnopyranosyl-( 1 →2) ]-α-L-arabinopyranosyl) oxy]-30-norolean-12,20( 29)-dien-28-oic acid O-ß-D-glucopyranosyl-( 1 → 6)-ß-D-glucopyranosyl ester( 2),3-O-ß-D-[( α-L-xylopyranosyl-( 1 → 2)-O-α-L-arabinopyranosyl)oxy]-30-norolean-12-en-28-oic acid α-L-rhamnopyranosyl-( 1 → 4)-O-ß-D-glucopyranosyl-( 1 → 6)-O-ß-D-glucopyranosyl ester( 3), yemuoside YM27( 4), yemuoside YM21( 5),yemuoside YM10( 6) and yemuoside YM7( 7). Conclusion: Compounds 1 ~ 3 are isolated from this plant for the first time.

[n-Butyl Alcohol-soluble Chemical Constituents of Psidium guajava Leaves].

OBJECTIVE: To study the chemical constituents of the leaves of Psidium guajava. METHODS: The chemical constituents were isolated by column chromatography on silica gel, Sephadex LH-20 and MPLC. Their chemical structures were elucidated on the basis of special analysis. RESULTS: Seven compounds were isolated from n-butyl alcohol fraction, whose structures were elucidated as morin-3-O-α-L-arabopyranoside (1), morin-3-O-α-L-iyxopyranoside (2), 2,6-dihydroxy-4-O-ß-D-glucopyranosyl-benzophenone (3), casuarictin (4),2,6-dihydroxy-3,5-dimethyl-4-O-(6"-O-galloyl-ß-D-glucopyranosyl)-benzophenone(5), globulusin A(6), and kaempferol-3-O-ß-D-(6"-galloyl) galactopyranoside (7). CONCLUSION: Compounds 3 and 5 ~ 7 are isolated from this plant for the first time.

Four New Triterpenoids from Callicarpa kwangtungensis.

Four new triterpenoids which were identifed as 2α,3ß,6ß,19α-tetrahydroxy- oleanolic acid 28-O-ß-D-glucopyranoside (1), 2-O-ß-D-glucopyranosyloxy-3α,19α-di-hydroxyoleanolic acid (2), 2-O-ß-D-glucopyranosyloxy-3α,19α-dihydroxyursolic acid (3), 2α,3α,6ß,19α-tetrahydroxyursolic acid 28-O-ß-D-glucopyranoside (4), were isolated from the aerial parts of Callicarpa kwangtungensis together with three known triterpenoids identified as 2α,3ß,21ß-trihydroxyursolic acid 28-O-ß-D-glucopyranoside (5), 2α,3α,19α,23-tetrahydroxyoleanolic acid 28-O-ß-D-glucopyranoside (6), 2α,3α,19α,23-tetrahydroxyursolic acid 28-O-ß-D-glucopyranoside (7). Their structures were elucidated by the combination of mass spectrometry (MS), one and two-dimensional NMR experiments.

Three new labdane-type diterpene glycosides from fruits of Rubus chingii and their cytotoxic activities against five humor cell lines.

Three new labdane-type diterpene glycosides, 15,18-di-O-ß-d-glucopyranosyl-13(E)-ent-labda-7(8),13(14)-diene-3ß,15,18-triol (1), 15,18-di-O-ß-d-glucopyranosyl-13(E)-ent-labda-8(9),13(14)-diene-3ß,15,18-triol (2), and 15-O-ß-d-apiofuranosyl-(1→2)-ß-d-glucopyranosyl-18-O-ß-d-glucopyranosyl-13(E)-ent-labda-8(9),13(14)-diene-3ß,15,18-triol (3), were isolated from the fruits of Rubus chingii. Their structures were elucidated on the basis of spectroscopic data and chemical methods. The cytotoxic activities of compounds 1-3 were evaluated against five human tumor cell lines (HCT-8, BGC-823, A549, and A2780). Compounds 3 showed cytotoxic activity against A549 with an IC50 value of 2.32µM.

[Chemical Constituents from Camellia oleifera Stem].

OBJECTIVE: To study the chemical constituents of stem of Camellia oleifera. METHODS: The chemical constituents were isolated and purified by column chromatography on silica gel, ODS, Sephadex LH-20 and MPLC. Their structures were elucidated on the basis of physicochemical properties and special analysis. RESULTS: Seven compounds were isolated from the stem of Camellia oleifera, whose structures were elucidated as (-) -pinoresinol (1), (-) -medioresinol (2), skullcapflavone II (3), betulinic acid (4), ursolic acid (5), 3-O-ß-D-glucopyranosyl- (1 --> 2) -ß-D-xylopyransoyl-(1 --> 3) -[ß-D-glucopyranosyl- (1 --> 2)] -ß-D-glucuronopyranosyl-22α-angeloyloxyolean-12-ene-15α,16α,28-triol (6) and oleanolic acid (7). CONCLUSION: Compounds 1 - 6 are isolated from this plant for the first time, and compounds 1 - 3 are isolated from this genus for the first time.

[Chemical Constituents from Callicarpa nudiflora].

OBJECTIVE: To study the chemical constituents of Callicarpa nudiflora. METHODS: The chemical constituents were isolated and purified by column chromatography on silica gel, ODS, Sephadex LH-20 and MPLC. Their structures were elucidated on the basis of physicochemical properties and special analysis. RESULTS: Eleven compounds were isolated from the leaves of Callicarpa nudiflora, whose structures were elucidated as 2α,3α-dihydroxyurs-12-en-28-oic acid (1), isorhamnetin (2), 2α,3ß,19α-trihydroxyurs-12-en-28-oic acid(3), 2α,3α,19α,23-tetrahydroxyurs-12-en-28-oic acid(4), 2α,3α,19α-trihyhydroxy-olean-12-en-28-O-α-D-glucopyranoside (5), benzyl-4'-hydroxy-benzoyl-3'-O-ß-D-glucopyranoside(6) (3S,5R,6R,7E,9S)-megastigman-7-ene-3,5,6,9-tetrol(7), philonotisflavone(8), 1, 6-di-O-caffeoyl-ß-D-glucopyranoside (9), luteolin-4'-O-(6"-E-caffeoyl)-ß-D-glucopyranoside (10), and (6S, 9R)-roseoside(11). CONCLUSION: All compounds are isolated from this plant for the first time.

Two new triterpenoids from Callicarpa kwangtungensis.

Two new triterpenoids, 2α,3ß,16α,19α,23-pentahydroxyolean-12-en-28-oic acid (1) and 2α,3α,11α,21α,23-pentahydroxyurs-12-en-28-oic acid (2), were isolated from the aerial parts of Callicarpa kwangtungensis. Their structures were elucidated by 1D and 2D analyses, as well as MS and IR spectra.

Hepatoprotective triterpenoid saponins from Callicarpa nudiflora.

Four new triterpenoid saponins, 2α,3α,19α,24-tetrahydroxyolean-12-en-28-oic-acid 28-O-ß-D-glucopyranosyl ester (1), 2α,3α,19α,23-tetrahydroxyolean-12-en-28-oic-acid 28-O-ß-D-xylopyranosyl-(1→2)-ß-D-glu-copyranosyl ester (2), 2α,3α,19α-trihydroxyolean-12-en-28-oic-acid 28-O-ß-D-xylopyranosyl-(1→2)-ß-D-glucopyranosyl ester (3), 2α,3α,23,29-tetrahydroxyurs-12,19-dien-28-oic-acid 28-O-ß-D-glucopyranosyl ester (4), together with three known compounds (5-7), were isolated from the leaves of Callicarpa nudiflora HOOK. Their structures were established by means of spectroscopic methods and chemical evidence. Hepatoprotective activities of the isolated compounds against D-galactosamine-induced toxicity have been tested. Among them, compounds 1-3 showed pronounced hepatoprotective activities against D-galactosamine-induced toxicity in WB-F344 rat hepatic epithelial stem-like cells.