Two new alkaloids from the bulbs of Lycoris sprengeri.

Two new alkaloids, lycosprenine (1) and 2α-methoxy-6-O-methyllycorenine (2), along with 22 known alkaloids (3-23b), were isolated from the bulb of Lycoris sprengeri. Their structures were elucidated on the basis of spectral analysis and by comparison of the spectroscopic data with those of known compounds. Selected compounds (1-3 and 6-9) were tested for their neuroprotective activities against H2O2-, CoCl2- and Aß25-35-induced SH-SY5Y cell injury, most of which exhibited neuroprotective effects of different degrees.

Three new ursane-type triterpenoids from the aerial parts of Isodon excisoides.

Three new ursane-type triterpenoids, 2α,3ß-dihydroxy-11α,12α-epoxy-urs-28,13ß-olide (1), 2α,3ß,24-trihydroxy-11α,12α-epoxy-urs-28,13ß-olide (2), and 2α,3α,24-trihydroxy-11,20(30)-dien-urs-28,13ß-olide (6), together with six known ursane-type triterpenoids (3-5, 7-9), were isolated from the EtOAc extract of the aerial parts of Isodon excisoides. Their structures were elucidated on the basis of 1D NMR and 2D NMR analyses as well as HRMS experiments.

Two new 5,8-quinoflavans from the leaf of Ilex centrochinensis.

Two new 5,8-quinoflavans were isolated from the leaf of Ilex centrochinensis, and their structures were elucidated as (2R)-7,3',4'-trimethoxy-5,8-quinoflavan and (2S)-7-methoxy-4'-hydroxy-5,8-quinoflavan on the basis of spectroscopic methods, especially 1D and 2D NMR, CD, and mass spectral analyses. Both of them exhibited weak cytotoxic activity against HuH7 cell lines and no cytotoxic activity against CaCO-2 cell lines.

Flavans from the leaf of Ilex centrochinensis.

Phytochemical investigation of the leaf of Ilex centrochinensis led to the isolation and characterization of four flavans, one flavanone (5), and one flavone (6), including a new compound whose structure was elucidated as (2S)-5,3',4'-trihydroxy-7-methoxyflavan (1) and a new natural product whose structure was elucidated as (2S)-5-hydroxy-7,3',4'-trimethoxyflavan (4) on the basis of spectroscopic methods especially 1D and 2D NMR, CD, and mass spectral analyses. Compounds 1 and 4 exhibited weak cytotoxic activity against Huh7 cell line and no cytotoxic activity against Caco-2 cell line.

Immunosuppressive constituents from Urtica dentata Hand.

A novel biscoumarin, 6,6',7,7'-tetramethoxyl-8,8'-biscoumarin (1), was isolated from the ethyl acetate extract of Urtica dentata Hand, together with five known compounds named as 7,7'-dihydroxy-6,6'-dimethoxy-8,8'-biscoumarin (2), 7,7'-dimethoxy-6,6'-biscoumarin (3), scoparone (4), vanillic acid (5), and daucosterol (6). Structures of the isolated compounds were elucidated on the basis of spectroscopic analysis including 2D NMR experiments. Compounds 1 and 2 were confirmed to be a rare carbon-carbon linked symmetrical biscoumarin. Compounds 1-4, especially 1 (IC(50) = 8.18 x 10(- 5) mol/l), showed potent immunosuppressive activities as determined by the Cell Counting Kit-8 assay for lymphocyte proliferation. Also, in the FACS analysis, 1 (IC(50) = 5.19 x 10(- 4) mol/l) promoted the differentiation of CD4(+)CD25(+)Foxp3(+) T regulatory cells distinctly compared to the normal control. Thus, 1 possessed specific immunosuppressive property by eliciting T regulatory cells, which may provide a potential treatment strategy for autoimmune diseases.

Anti-inflammatory Activity of Mollugin on DSS-induced Colitis in Mice.

We aimed to explore the anti-inflammatory activity of mollugin extracted from Rubia cordifolia L, a traditional Chinese medicine, on dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) in mice. Thirty C57BL/6 mice were divided into a control group (n=6), a model group (n=6), and three experimental groups (40, 20, 10 mg/kg of mollugin, n=6 each). DSS solution (3%) was given to mice in the model group and experimental groups from day 4 to day 10 to induce the mouse UC model. Mice in the experimental groups were intragastrically administrated mollugin from day 1 to day 10. Animals were orally given distilled water in the control group for the whole experiment time and in the model group from day 1 to day 3. The changes in colon pathology were detected by hematoxylin and eosin (HE) staining. Interleukin-1ß (IL-1ß) in the serum, and tumor necrosis factor-α (TNF-α) and interferon-γ (IFN) in the tissues were measured by enzyme linked immunosorbent assay. Expression levels of Toll-like receptor 4 (TLR4) and myeloid differentiation factor 88 in the colon tissues were detected by immunohistochemistry. Results showed that mollugin could significantly reduce weight loss and the disease activity index in the DSS-induced UC mouse model. HE examinations demonstrated that mollugin treatment effectively improved the histological damage (P<0.05). The overproduction of IL-1ß and TNF-α was remarkably inhibited by mollugin treatment at doses of 20 and 40 mg/kg (P<0.05). Additionally, the levels of TLR4 in colon tissues were significantly reduced in mollugin-treated groups compared with the DSS group. Our findings demonstrated that mollugin ameliorates DSS-induced UC by inhibiting the production of pro-inflammatory chemocytokines.

Structural analysis and antitussive evaluation of five novel esters of verticinone and bile acids.

Shedan-Chuanbei powder, a complex of traditional Chinese medicine preparation, which consists of Snake Bile (Chinese name "Shedan") and Fritillariae Cirrhosae (Chinese name "Chuanbei"), is the most popular antitussive and expectorant formulation in Chinese communities. However, the clinical application of Shedan-Chuanbei powder is now stringently limited because of the shortage of the two crude medicinal materials, especially for the sake of animal protection. In addition, the inherent defects of the most of the complex of traditional Chinese medicine such as the indistinct basal pharmacodynamic materials and the difficulties in quality control had blocked them heading into the international medicinal market. So we attempted to seek new substitute for Shedan-Chuanbei powder for antitussive drugs. In order to gain some new compounds with better bioactivity and attenuated toxicity, we tried to combine two kinds of drugs through ester bond. Enlightened with "combination principle" in drug discovery, we synthesized five novel esters of verticinone and bile acids, both of which are the major bioactive components in Shedan-Chuanbei powder. We then evaluated the antitussive activity and the acute toxicity of the five ester-linked compounds. The five ester-linked compounds had much more potent antitussive activity and expectorant activity than single bile acids at the same doses, and had equivalent antitussive activity and expectorant activity in comparison with about double moles dose of the monomer verticinone. Especially, cholic acid-verticinone ester had much more potent antitussive effects than the monomer verticinone or cholic acid at the same dose. A further acute toxicity study showed that the LD(50) values of the five ester-linked compounds exceeded 3.5g/kg by intraperitoneal injection in mice. Based on the studies of pharmacology and acute toxicity, the five ester-linked compounds have synergic pharmacodynamic action and attenuated toxicity compared with single verticinone and single bile acids.

Addictive evaluation of cholic acid-verticinone ester, a potential cough therapeutic agent with agonist action of opioid receptor.

AIM: The purpose of this work was to search for potential drugs with potent antitussive and expectorant activities as well as a low toxicity, but without addictive properties. Cholic acid-verticinone ester (CA-Ver) was synthesized based on the clearly elucidated antitussive and expectorant activities of verticinone in bulbs of Fritillaria and different bile acids in Snake Bile. In our previous study, CA-Ver showed a much more potent activity than codeine phosphate. This study was carried out to investigate the central antitussive mechanism and the addictive evaluation of CA-Ver. METHODS: Testing on a capsaicin-induced cough model of mice pretreated with naloxone, a non-selective opioid receptor antagonist, was performed for the observation of CA-Ver's central antitussive mechanism. We then took naloxone-induced withdrawal tests of mice for the judgment of CA-Ver's addiction. Lastly, we determined the opioid dependence of CA-Ver in the guinea pig ileum. RESULTS: The test on the capsaicin-induced cough model showed that naloxone could block the antitussive effect of CA-Ver, suggesting the antitussive mechanism of CA-Ver was related to the central opioid receptors. The naloxone-urged withdrawal tests of the mice showed that CA-Ver was not addictive, and the test of the opioid dependence in the guinea pig ileum showed that CA-Ver had no withdrawal response. CONCLUSION: These findings suggested that CA-Ver deserved attention for its potent antitussive effects related to the central opioid receptors, but without addiction, and had a good development perspective.

Triterpenoids from the roots of Actinidia chinensis.

Two new triterpenoids, 1 and 2, were isolated from the hepatoprotective AcOEt fraction of the roots of Actinidia chinensis, together with eight known 12-en-28-oic acids of oleanane or ursane type, 3-10. The two new compounds were elucidated as 2alpha,3beta-dihydroxyurs-12-en-28,30-olide (1) and 2alpha,3beta,24-trihydroxyurs-12-en-28,30-olide (2), on the basis of spectroscopic (IR, NMR, and MS) analyses. The chemotaxonomic significances of some triterpenoids were also discussed.

Two new triterpenoids from the leaves and stems of Fritillaria hupehensis.

Two new cycloartane-type triterpenoids 25-hydroxyl-9,19-cycloart-22-ene-3-one (1) and (23Z)-9,19-cycloart-23-ene-3alpha,25-diol (2) along with 9,19-cycloart-25-ene-3beta, 24xi-diol (3) and cycloeucalenol (4) have been isolated from the leaves and stems of Fritillaria hupehensis Hsiao et K.C. Hsia. Their structures were elucidated on the basis of spectroscopic analysis.

The interleukin-18 inhibitory activities of echinocystic acid and its saponins from Impatiens pritzellii var. hupehensis.

Echinocystic acid (1), an echinocystic acid saponin, 2, and four of its ester saponins, 3-6, obtained from the active fraction of Impatiens pritzellii var. hupehensis, an traditional Chinese medicine for rheumatoid arthritis, were investigated for their effects on lipopolysaccharide (LPS)-induced interleukin (IL)-18 in human peripheral blood mononuclear cells. Three of them, 1, 2 and 6, showed obvious activity to inhibit the production of IL-18, especially the ester saponins with a sugar chain at C-28, 6. Structure-activity relationships are discussed in brief.

[Studies on the chemical constituents of Sarcopyramis nepalensis].

OBJECTIVE: To study the chemical constituents of Sarcopyramis nepalensis. METHODS: The constituents were isolated and purified with chromatography and the structures were elucidated by spectral analysis. RESULTS: Ten compounds were isolated and their structures were identified as: (E)-1-(3,4-dihydroxy-phenyl) ethyl acrylate (I), stearic acid (II), palmitic acid (III), 4-hydroxybenzonic acid (IV), 3,4-dihydroxy benzoic acid (V), gentisic acid ( VI), gallic acid (VII), beta-sitosterol (VIII), daucosterol (IX), stigmasterolstearate (X). CONCLUSION: Compounds I , IV, V, VI, VII are isolated from this plant for the first time.

Cytotoxic alkaloids from the bulbs of Fritillaria hupehensis.

Phytochemical investigation of the bulbs of Fritillaria hupehensis resulted in the isolation and structural elucidation of four new steroidal penta- and hexacyclic veratraman- and cevan-based alkaloids, respectively, compounds 1-4. They were obtained together with the known constituents ebeinine (5) and zhebeinine (6), which were isolated for the first time from this plant. The structures of the new isolates were established by spectroscopic and mass-spectrometric analyses, in combination with chemical methods. All compounds were assayed for their cytotoxic effects towards HeLa and HepG2 cell lines. Compounds 1 and 2 showed significant inhibitory effects against both types of tumor cells, with IC(50) values in the range 2.52-0.23 microM, similar as those for 5-fluorouracil used as positive control.

[Studies on chemical constituents from Anoectochilus roxburghii].

OBJECTIVE: To investigate the chemical constituents from Anoectochilus roxburghii. METHODS: The compounds were isolated and purified by repeated column chromatographies with silica gel, Macroporous resin and Sephadex LH-20, and their structures were identified by their physical and spectral datas. RESULTS: Ten compounds were isolated and elucidated as: beta-D-glucopyranosyl-(3R)-hydroxybutanolide (I), stearic acid (II), palmitic acid (III), beta-sitosterol (IV) and succinic acid (V), p-hydroxy benzaldehyde (VI), daucosterol (VII), and methyl 4-beta-D-glucopyranosyl-hutanoate (VIII); as well as p-hydroxy cinnamic acid (IX) and o-hydroxy phenol (X) were identified. CONCLUSION: Compound I ,VIII, X are firstly isolated from this species.

Two New Ursane-type Triterpenoid Glycosides from Ilex Cornuta.

Ilex cornuta (I. cornuta) is a traditional Chinese medicine (TCM) that has been used in clinical practice for hundreds of years. In order to provide more information about the chemical basis of its pharmacological effects, phytochemical investigation on the roots of I. cornuta was conducted in this study. The roots of the plant were firstly extracted with 95% EtOH, and then the crude was partitioned with petroleum ether, EtOAc and n-butyl alcohol. Different chromatographies were employed to isolate the crude step by step and the crude was further purified by semipreparative high performance liquid chromatography (HPLC). As a result, two new triterpenoid saponins (1, 2), together with 12 known compounds (3-14), were isolated from the roots of I. cornuta. Their structures were determined based on nuclear magnetic resonance (NMR), mass spectrum (MS) technologies, chemical reactions as well as gas chromatography (GC). Compounds 4, 6, 8, 11, 12 and 13 were isolated from this genus for the first time. The structures of compounds 1 and 2 were determined as 3ß-O-α-D-xylopyranosyl-(1→3)-α-L-2-O-acetylarabinopyranosyl-(1→2)-ß-D glucopyranosyl-23-hydroxyl-20α(H)-urs-12-en-28-oic acid 28-O-ß-D-glucopyranosyl ester (1) and 3ß-O-α-D-xylopyranosly-(1→3)-α-L-2-O-acetylarabinopyranosyl-(1→2)-ß-Dglucopyranosyl- 19α,23-dihydroxyl-20α(H)-urs-12-en-28-oic acid 28-O-ß-D-glucopyranosyl ester (2).

Ferulic acid esters from Euphorbia hylonoma.

Octacosyl cis-ferulate (1), along with the trans isomer (2), cholest-5-en-3beta-ylhexadecanoate, chrysophanol and octadecanoic acid was isolated from the roots of Euphorbia hylonoma.

[Preparation and antitussive, expectorant and antiasthmatic activities of verticinone-bile acids salts].

To search for potential drugs with potent antitussive, expectorant, antiasthmatic activities and low toxicity, a series of verticinone-bile acids salts were prepared based on the clearly elucidated antitussive, expectorant and antiasthmatic activities of verticinone in bulbs of Fritillaria and different bile acids in Snake Bile. The antitussive, expectorant and antiasthmatic activities of these verticinone-bile acid salts were then screened with different animal models. Ver-CA (verticinone-cholic acid salt) and Ver-CDCA (verticinone-chenodeoxycholic acid salt) showed much more potent activities than other compounds. The bioactivities of Ver-CA and Ver-CDCA are worthy to be intensively studied, and it is also deserved to pay much attention to their much more potent antitussive effects than codeine phosphate. In order to elucidate whether they have synergistic effect and attenuated toxicity, their activities will be continuously compared with single verticinone, cholic acid and chenodeoxycholic acid at the same doses on different animal models. The application of "combination principles" in traditional Chinese medicinal formulations may be a novel way in triditional Chinese medicine research and discovery.

[Studies on constituents from roots of Euphorbia hylonoma].

OBJECTIVE: To study the constituents from roots of Euphorbia hylonoma. METHOD: Column chromatographic techniques were used for isolation and purification of the chemical constituents and their structures were identified by spectral analysis (IR, 1H-NMR, 13C-NMR, 2D-NMR and MS). RESULT: Six compounds were isolated and elucidated as nonane (1), bis (2-ethylhexyl) phthalate (2), euphol (3), beta-sitosterol (4), acalyphol (5) and daucosterol (6) respectively. CONCLUSION: Compounds 1, 2, 3, 5 and 6 were isolated from the plant for the first time.

[HPLC fingerprint of Fritillaria hupehensis].

AIM: To establish a fingerprint analysis method of Fritillaria hupehensis. METHODS: fingerprint was performed by HPLC-ELSD. Hypersil ODS column was used; the mobile phase was composed of methanol (with 0.05% triethylamine) and water with gradient elution; flow rate was 1.0 mL x min(-1); recording time was 60 min; drift tube temperature was 75 degrees C; gas flow rate was 1.9 L x min(-1). RESULTS: HPLC fingerprint of Fritillaria hupehensis was obtained. CONCLUSION: A reliable method was provided for controlling the quality of Fritillaria hupehensis.

Alkaloids from the bulbs of Lycoris longituba and their neuroprotective and acetylcholinesterase inhibitory activities.

Three novel alkaloids (1-3), together with nineteen known ones (4-22), were isolated from the bulbs of Lycoris longituba. Their structures were elucidated on the basis of extensive spectroscopic analyses, which belong to several Amaryllidaceae alkaloid skeletons. Among them, the harmane-type alkaloids (the new compound 1 and the known compounds 5, 6 and 7) were found for the first time from Lycoris genus. The isolates were tested for their neuroprotective activities against CoCl2, H2O2 and Aß25-35-induced SH-SY5Y cell injuries, and the majority of them exhibited neuroprotective activities of different degrees. The acetylcholinesterase (AChE) inhibitory activities of the isolated alkaloids were also evaluated, while compounds 12, 14-20 and 22 exhibited extremely significant AChE inhibitory activities.