Inhibitory effect of tanshinones on rat CYP3A2 and CYP2C11 activity and its structure-activity relationship

This study investigated the effect of tanshinones on rat liver microsomal CYP3A2 and 2C11 activity and explored the structure-activity relationship of tanshinones with CYP3A activity. Cryptotanshinone, tanshinone I and tanshinone IIA were competitive CYP3A2 inhibitors (K_i = 199-243 μM) and CYP2C11 inhibitors (K_i = 91-118 μM). Dihydrotanshinone was not only a noncompetitive inhibitor of CYP3A2 (K_i = 110 μM), but also a competitive CYP2C11 inhibitor (K_i = 55 μM). The structural difference between dihydrotanshinone and tanshinone I at C-15 position of furan ring resulted in the different modes of inhibition on CYP3A activity.     

Anti-thrombotic activity and chemical characterization of steroidal saponins from Dioscorea zingiberensis C.H. Wright

Steroidal saponins have long attracted scientific attention, due to their structural diversity and significant biological activities. Total steroidal saponins (TSS) extracted from the rhizomes of Dioscorea zingiberensis C.H. Wright (DZW) constitute an effective treatment for cardiovascular disease. However, the active constituents contained in DZW rhizomes and their pharmacological properties are not fully understood. The aim of this work is to determine and quantify the active constituents in DZW rhizomes using fingerprint technique, and evaluate its anti-thrombotic activity using inferior vena cava ligation thrombosis rat model and pulmonary thrombosis mice model after being gavaged with TSS for 1 or 2 weeks. In the study, a chemical fingerprint method was firstly established and validated to quantify and standardize TSS from DZW rhizomes including parvifloside, protodeltonin, protodioscin, protogracillin, zingiberensis saponin, deltonin, dioscin and trillin. TSS extracted from DZW rhizomes were showed to have the inhibitions on platelet aggregation (PAG) and thrombosis, and prolong activated partial thromboplastin time (APTT), thrombin time (TT), and prothrombin time (PT) in a dose-dependent manner in rats. TSS also prolonged the bleeding time and clotting time in a dose-dependent manner in mice. The results indicate that TSS could inhibit thrombosis by both improving the anticoagulation activity and inhibiting PAG action, suggesting that TSS from DZW rhizomes have the potential to reduce the risk of cardiovascular diseases by anti-thrombotic action.     

A new abietane diterpene from Glyptostrobus pensilis

A new abietane diterpene, glypensin A (1) and four known compounds, 12-acetoxy-ent-labda-8(17), 13E-dien-15-oic acid (2), quercetin 3-O-α-L-arabinofuranoside (3), quercetin 3-O-β-D-galactopyranoside (4), β-sitosterol (5) were isolated from the branches and leaves of Glyptostrobus pensilis (Staut.) Koch. Their structures were determined by MS, 1D- and 2D-NMR means. Compound 1 showed cytotoxicity on human chronic myeloid leukemia cell line K562 (IC₅₀ = 21.2 μM).     

Isolation, stability and bioactivity of Jatropha curcas phorbol esters

Jatropha curcas seed oil, which can be utilized for biodiesel production upon transesterification, is also rich in phorbol esters (PEs). In this study, PEs from J. curcas oil (Jatropha factors C₁ and C₂ (purified to homogeneity), Jatropha factors C₃ and (C₄ + C₅) (obtained as mixtures) and PE-rich extract (containing all the above stated Jatropha factors) were investigated. The concentrations of Jatropha PEs were expressed equivalent to Jatropha factor C₁. In the snail (Physa fontinalis) bioassay, the order of potency (EC₅₀, μg/L) was: PE-rich extract < factor C₃ mixture < factor C₂ < factor C₁ < factor (C₄ + C₅). In the Artemia salina bioassay, the order of potency (EC₅₀, mg/L) was: factor C₂ < factor C₃ mixture < factor C₁ < factor (C₄ + C₅) mixture. In addition, Jatropha PEs exhibited platelet aggregation (ED₅₀, μM, factor C₂ < factor C₃ mixture < factor C₁ < factor (C₄ + C₅) mixture. The stability of a PE-rich extract was evaluated and found to be low at room temperature but favourable in ethanol over a range of temperatures. By integrating the isolation methodology developed in this study in the Jatropha biodiesel industry, PEs could be obtained as value-added co-products.     

The role of seasonality on the inhibitory effect of Brazilian green propolis on the oxidative metabolism of neutrophils

The reactive oxygen species (ROS) produced by neutrophils are involved in the pathogenesis of several diseases, for which the intake of antioxidants could benefit patients either as a prophylactic or therapeutic treatment. Propolis is among the known antioxidants, and its chemical composition may vary under the influence of seasonality, which may interfere in its biological properties. This work evaluates the role of seasonality on the production of some important compounds of propolis samples produced monthly from November 2001 through October 2002 as well as the effect of these samples on the oxidative metabolism of stimulated neutrophils, by using both luminol and lucigenin to produce chemiluminescence (CLlum and CLluc, respectively). The cytotoxicity of the most active extracts to neutrophils was also investigated. The inhibitory effect of the propolis samples varied significantly during the studied period for both assays (3.4 ± 1.1 to 16.0 ± 1.1 μg/mL for CLlum and 6.2 ± 2.0 to 30.0 ± 5.0 μg/mL for CLluc), which was also observed in the quantitative profile of the main analyzed compounds (aromadendrin-4′-methyl ether, artepillin C, and baccharin). This effect started to become more prominent during the fall and, among all the studied extracts, the one obtained in May displayed the highest inhibitory effect on CL production (3.4 ± 1.1 μg/mL for CLlum and 6.2 ± 2.0 μg/mL for CLluc). The HPLC qualitative profiles of the extracts of propolis samples were quite similar, but there was a huge variation in terms of quantitative profile. It seems that aromadendrin-4′-methyl ether and baccharin play an essential role in the antioxidant activity, while artepillin C is not very important for this effect. The extracts presenting the highest antioxidant activity were produced in May, June, and August, and they did not display cytotoxicity at 25 μg/mL; quercetin, used as control, was not toxic to neutrophils at 8.5 μg/mL.     

Structural insights to investigate Conypododiol as a dual cholinesterase inhibitor from Asparagus adscendens

The main aim of the current study was to explore molecular insights for potentially new dual cholinesterase inhibitor(s) from Asparagus adscendens via molecular docking. This medicinal plant is traditionally used as a nerve tonic and remedy for memory impairments. Conypododiol was isolated from the chloroform fraction of methanolic extract of A. adscendens, based on bioactivity guided isolation. Conypododiol exhibited significant inhibition of both acetylcholinesterase and butyrylcholinesterase, having the IC₅₀ values 2.17 ± 0.1 μM and 11.21 ± 0.1 μM, respectively. IC₅₀ values of the standard compound Galanthamine for both the enzymes were 0.537 ± 0.018 μM and 8.6 ± 0.27 μM, respectively. Based on MTT cytotoxicity assay, Conypododiol was found safe against LCMK-2 monkey kidney epithelial cells and mice hepatocytes. Molecular docking studies revealed the hydrogen bonding interactions of Conypododiol with His440 and Ser200 at esteratic site (ES), and also with Tyr334 at peripheral anionic site (PAS) of the aromatic gorge of acetylcholinesterase. Simultaneous contacts of Conypododiol with PAS and ES shows its significance as a bivalent ligand. This preliminary study highlighted the potential of Conypododiol to be further developed and modified as new lead compound identified by its folk use.     

Two new amide alkaloids with anti-leukaemia activities from Aconitum taipeicum

Two new amide alkaloids, named 3-isopropyl-tetrahydropyrrolo [1,2-a] pyrimidine-2,4(1H,3H)-dione (1) and 1-acetyl-2,3,6-triisopropyl-tetrahydropyrimidin-4(1H)-one (2), were isolated from the roots of Aconitum taipeicum. Their chemical structures were characterized on the basis of MS, 1D- and 2D NMR. The anti-leukaemia activities of the compounds were also tested. The results indicated that the compounds exhibited more significant cell growth inhibitory activities against HL-60 cells than adriamycin, with the IC₅₀ of 1.1 ± 0.03 μg/mL and 1.6 ± 0.07 μg/mL respectively. In addition, two compounds showed anti-tumor activities against K562 cells as well.     

Antimicrobial activity of bioactive compounds and leaf extracts in Jatropha tanjorensis

Objectives

Jatropha tanjorensis was investigated scientifically to generate evidence for the efficacies reported in traditional systems and the results are given here.

Methods

Different concentrations of the solvent extracts of leaves and four isolated compounds were tested against human pathogenic microorganisms such as gram-positive bacteria of Bacillus cereus, Bacillus subtilis, Staphylococcus aureus, Staphylococcus epidermis, gram-negative bacteria of Aeromonas hydrophila, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus mirabilis, Proteus vulgaris, Salmonella paratyphi, Salmonella paratyphi A, Vibrio alcaligenes, Vibrio cholerae and fungi of Aspergillus fumigatus, Candida albicans, Microsporum gypseum and Trichophyton rubrum by agar-well diffusion and disk diffusion methods.

Results

In agar-well diffusion method, maximum activity was recorded in a concentration-dependent manner. The extracts recorded activity against bacteria such as 17–26, 15–25 and 13–24 mm to methanol extract and 15–24, 14–23 and 12–22 mm to chloroform extract at 50, 25 and 12.5 mg/ml respectively and fungi such as 9–15 mm to A. fumigatus and 5–16 mm to T. rubrum. Maximum activity was 30–46, 27–43 and 17–40 mm to friedelin and 23–46, 28–44 and 18–41 mm to R (+) 4-hydroxy-2-pyrrolidinone against bacteria and 12–37, 8–34 and 31–33 mm to friedelin and 12–40, 11–35 and 10–33 mm to R (+) 4-hydroxy-2-pyrrolidinone against fungi at 10, 5 and 2.5 mg respectively.

Conclusions

The present study concludes that friedelin, β-amyrin, stigmasterol and R (+) 4-hydroxy-2-pyrrolidinone present in the methanol extract could be responsible for the broad spectrum of antimicrobial activity and provide scientific evidence.     

Comparative characteristic of the inflammatory diterpenes in the roots of Euphorbia fischeriana with different preparation method using HPLC-ELSD

A rapid and simple method was established for the simultaneous determination of ten diterpenes by reversed phase HPLC coupled with evaporative light scattering detection. Chromatographic separation was carried out in gradient mode by using a WondaSil™ C₁₈ column (250 mm × 4.6 mm, 5 μm) with mobile phases of methanol and water at 1 mL/min. The drift tube temperature of evaporative light scattering detector was set to 65 °C and nitrogen flow-rate was 2.7 L/min. The method validated was shown to be specific, precise, accurate and linear. Moreover, it was applied to investigate four samples of E. fischeriana with different extracting methods. Contrast to the dried roots, the fresh roots had much higher content of prostratin which represented much higher inflammatory effects than other diterpenes. The results demonstrated that the dried roots were suitable for the ordinary therapy to avoid intense stimulatory, while the fresh roots could be used in the anticancer treatment. All of the results suggested that comparative analysis of chemical components as biomarker and connecting toxic effects of an herb was helpful for revealing the mechanism of its toxicity, and for guiding safer and better application of the herb in TCM clinic.     

The novel Sinupret® dry extract exhibits anti-inflammatory effectiveness in vivo

Sinupret® is frequently used as a herbal medicinal product to treat sinusitis, and it was assumed that anti-inflammatory effects might contribute to its overall beneficial properties. Here, we investigated the effects of a Sinupret® drug mixture (SIN) as well as of the novel Sinupret® dry extract (SIN DE) with the latter containing higher concentrations of active ingredients, in an in vivo model of acute inflammation, the carrageenan-induced pleurisy in rats. Both SIN and SIN DE were administered to rats orally at doses of 100 mg/kg (low dose) and 500 mg/kg (high dose) 1 h prior to intrapleural injection of carrageenan. Although both SIN and SIN DE significantly reduced the exudate volume and leukocyte numbers in the pleural exudate at the high and the low dose 4 h after carrageenan injection, the novel SIN DE was more efficient than SIN at the low dose, implying higher efficiency. In parallel, the novel dry extract SIN DE, but not SIN, at 500 mg/kg significantly lowered the levels of prostaglandin (PG)E₂ in the exudates and reduced the amounts of cyclooxygenase (COX)-2 protein in the lungs. Together, SIN and SIN DE exert significant oral anti-inflammatory effects, which rationalize their therapeutic use in the management of sinusitis and other viral/microbial nasal infections that are associated with inflammation. Moreover, our results suggest that based on the higher efficiency and the accompanied reduction of COX-2 expression and PGE₂ formation, the novel dry extract SIN DE might be superior over the former SIN drug mixture.     

Cytotoxic taraxerane triterpenoids from Saussurea graminea

Four new taraxerane triterpenoids, 1β,3β-dihydroxy-11α,12α-oxidotaraxerane (1), 28-hydroxy-11α,12α-oxidotaraxerane-3-one (2), 3β-hydroxy-11α,12α-oxidotaraxerane-28-al (3), and 3-O-acetyl-11α,12α-oxidotaraxerane-28-al (4), together with three known compounds 3β-hydroxy-11α,12α-oxidotaraxerane (5), 3β,28-dihydroxy-11α,12α-oxidotaraxerane (6), and 11α,12α-oxidotaraxerane-3-one (7), were isolated from the 70% EtOH extract of Saussurea graminea. Their structures were elucidated on the basis of extensive 1D and 2D NMR (COSY, HMQC, HMBC and NOESY) analyses. The isolated compounds were evaluated in vitro for cytotoxic properties against eight tumor cell lines (A-549, BGC-823, HCT15, HeLa, HepG2, MCF-7, SGC-7901 and SK-MEL-2).     

Profiling of volatile compounds from five interior decoration timbers in Taiwan using TD/GC–MS/FID

Volatile organic compounds (VOCs) released from Chamaecyparis formosensis, Cryptomeria japonica, Cunninghamia lanceolata, Chamaecyparis obtusa var. formosana, and Taiwania cryptomerioides five major building and interior decoration timbers and their essential oil components were analyzed using GC–MS and TD/GC–MS/FID. Results showed that C. obtusa var. formosana had the highest yield of essential oil (3.42%), followed by C. formosensis (3.14%), while C. japonica had the lowest yield (0.95%). Moreover, oxygenated sesquiterpene was the highest relative content in all five essential oils and their main constituents were trans-myrtanol (18.04%), 1-epi-cubenol (15.99%), cedrol (62.26%), α-cadinol (26.42%), and α-cadinol (27.98%), respectively. In terms of emission quantity of top VOC, the results showed the decreasing order of C. formosensis (myrtenal, 74.21 mg/m²)?>?T. cryptomerioides (thujopsene, 12.00 mg/m²)?>?C. lanceolata (α-cedrene, 10.27 mg/m²)?>?C. obtusa var. formosana (α-pinene, 8.05 mg/m²)?>?C. japonica (α-cedrene, 4.25 mg/m²). C. formosensis had a greater amount of VOCs emitted and hence gave off more fragrance than C. obtusa var. formosana initially. However, after indoor exposure of 24 weeks, the VOC emission quantity of C. obtusa var. formosana exceeded that of C. formosensis. α-Cedrene and thujopsene were the top two major VOCs of both C. lanceolata and T. cryptomerioides. However, they both showed a trend of decrease in emission with prolonged exposure. All five plantation timbers showed good antifungal, antimicrobial, antibacterial, and antitermitic properties, making them ideal materials for interior decoration. Not only do they have strong bioactivities, they can also provide a fragrant and healthy living environment.     

A new hepoprotective saponin from Semen Celosia cristatae

A new triterpenoid saponin, named semenoside A (1), was isolated from Semen Celosia cristatae. Its structure was elucidated on the basis of 1D, 2D NMR, HR-FAB-MS and ESI-MS techniques, and physicochemical properties. The hepatoprotective activity of semenoside A with an oral dose of 1.0, 2.0 and 4.0 mg/kg, respectively, were investigated by carbon tetrachloride (CCl₄)-induced hepatotoxicity in mice. The results indicated that it had significant hepatoprotective effects (p < 0.01).     

Kujigamberol, a new dinorlabdane diterpenoid isolated from 85million years old Kuji amber using a biotechnological assay

A new compound, 15,20-dinor-5,7,9-labdatriene-18-ol (1), named kujigamberol, was isolated from amber, fossilized tree resin from the Kuji area in Japan, has been dated as being 85 million years old (late Cretaceous). Kujigamberol was identified using the hypersensitive mutant yeast (zds1? erg3? pdr1? pdr3?) with respect to Ca²⁺-signal transduction. The structure was elucidated on the basis of spectroscopic analysis including 1D NMR, 2D NMR and HR-EI-MS. It was different from known diterpenoids with a similar activity isolated from Baltic amber (agathic acid 15-monomethyl ester (2), dehydroabietic acid (3) and pimaric acid (4)). Kujigamberol showed glycogen synthase kinase-3β (GSK-3β) inhibition activity involving the growth restored activity against the mutant yeast and was cytotoxic to HL60 cells (IC₅₀ = 19.6 μM).     

Transdermal behaviors comparisons among Evodia rutaecarpa extracts with different purity of evodiamine and rutaecarpine and the effect of topical formulation in vivo

Aim

Evodiamine (EVO) and rutaecapine (RUT), the major active components from Evodia rutaecarpa extract (EE), are recognized as a depended analgesic agent. This study was designed to investigate the effect of purity and chemical enhancers on the transdermal behavior of EVO and RUT, and the pharmacological effect of their topical cream in vivo.

Material and methods

Transdermal delivery across a full thickness pig abdominal skin was detected in vitro by Franz-type diffusion cell, with HPLC for quantification of the permeation of EVO and RUT. The activity of topical cream in vivo was evaluated by a mice pain model induced by formalin and hot plate.

Results

Transdermal characters of EVO and RUT showed a low transdermal rate, long lag time and low cumulative amount. The transdermal rate and cumulative amount could be promoted by lipophilic enhancers, whereas lag time was shortened by hydrophilic surfactant, but these permeation parameters were not markedly influenced by purity of EE (p > 0.05). The effect in vivo was confirmed by analgesic models in topical cream of EE, which produced a significant (p < 0.05) inhibitory effects on pain response in dose-dependent manner.

Conclusion

The purity of EVO and RUT from EE has no significant effect on their permeation through porcine skin, but oleic acid or nerolidol can markedly elevate the transdermal rate of EVO and RUT. High purity of EE is the best choice for topical preparation to increase the drug loading. The effect of EE in vivo is verified by formalin model and hot plate test.     

Identification of non-alkaloid acetylcholinesterase inhibitors from Ferulago campestris (Besser) Grecescu (Apiaceae)

Inhibition of Acetylcholinesterase (AChE) is still considered as a strategy for the treatment of neurological disorders such as Alzheimer's disease. Many plant derived alkaloids (such as galantamine and rivastigmine) are known for their AChE inhibitory activity. Recently, other classes of natural compounds such as terpenoids, sesquiterpene glycosides and coumarins have been studied as new AChE inhibitors, with the aim to discover less toxic compounds compared to alkaloidal ones. The Ferulago campestris roots dichloromethane extract was used for a bioassay-guided fractionation for the search of AChE inhibitors. Three coumarin derivatives (umbelliprenin 1, coladonin 2 and coladin 3), three daucane ester derivatives (siol anisate 4, ferutinin 5 and 1-acetyl-5-angeloyl lapiferol 6), two phenol derivatives (2-epilaserine 7 and epielmanticine 8) and one polyacetylene (9-epoxyfalcarindiol 9) were isolated by the bioassay-guided approach. Their structures were characterized on the basis of spectral methods (1D and 2D NMR, and MS spectroscopy). All the isolated compounds were able to inhibit the AChE (IC₅₀ 1.2–0.1 mM) although at higher doses if compared to galantamine (6.7 μM) measured in the same conditions. The most active compounds were the daucane derivative siol anisate 4 and the epielmanticine 8, with IC₅₀ of 0.172 and 0.175 mM respectively.     

Alkaloids from Lycoris aurea and their cytotoxicities against the head and neck squamous cell carcinoma

Phytochemical investigation of the 80% EtOH extract of the bulbs of Lycoris aurea led to the isolation of six new alkaloids, 2-demethyl-isocorydione (1), 8-demethyl-dehydrocrebanine (2), 1-hydroxy-anhydrolycorin-7-one (3), (+)-1,2-dihydroxy-anhydrolycorine N-oxide (4), 5,6-dihydro-5-methyl-2-hydroxyphenanthridine (5), and (+)-8-hydroxy-homolycorine-α-N-oxide (6), and together with two known compounds, isocorydione (7) and anhydrolycorin-7-one (8). Structural elucidation of all the compounds was performed by spectral methods such as 1D and 2D (¹H–¹H COSY, HMQC, and HMBC) NMR spectroscopy, in addition to high resolution mass spectrometry. All the alkaloids were in vitro evaluated for their cytotoxic activities against seven tumor cell lines of the head and neck squamous cell carcinoma and anti-inflammatory activities. Compounds 1, 2, 6, and 7 exhibited significant cytotoxicities against all the tested cell lines. Moreover, alkaloids 1, 2, and 7 possessed selective inhibition of Cox-2 (> 85%).     

Furanocoumarins-imperatorin inhibits myocardial hypertrophy both in vitro and in vivo

The furanocoumarin imperatorin has been reported to have hypotensive effect, and we have investigated its activity on myocardial hypertrophy. Imperatorin displayed similar chromatographic retention peak to verapamil in the model of cardiac muscle/cell membrane chromatography, and could reduce in a concentration-dependent way protein content and cell size of myocytes prestimulated with angiotensin II. The ratio of heart weight to body weight (mg/g) (4.13 ± 0.06 in SHRs, 3.77 ± 0.02 with imperatorin 25 mg kg^− 1 d^− 1 16 17 ig, P < 0.01) was significantly lower in the imperatorin-treated SHRs. Taken together, our observations show that imperatorin exerts anti-hypertrophic effect both in vitro and in vivo.