Sesquiterpenes from the rhizomes of Alpinia japonica and their inhibitory effects on nitric oxide production

A novel norsesquiterpene (1), three new bisabolenes sesquiterpenes (2–4), along with 7 known compounds (5–11), were isolated from the rhizomes of Alpinia japonica. The structures of the new compounds were elucidated by analysis of spectroscopic data. The absolute configuration of C-1 in 3 was deduced via the circular dichroism data of the in situ formed [Rh₂(OCOCF₃)₄] complexes. Inhibitory effects of the isolates on nitric oxide production in lipopolysaccaride-activated RAW264.7 macrophages were evaluated. Compound 6 showed significant inhibitory activity with IC₅₀ value of 5.3 μΜ, and compounds 1, 3, 5 and 7–10 exhibited moderate inhibitory activities with IC₅₀ values between 24.5 and 46.3 μM.     

Indole alkaloids from the roots of Isatis indigotica and their inhibitory effects on nitric oxide production

Three rare indole-2-S-glycosides, indole-3-acetonitrile-2-S-β-D-glucopyranoside (1), indole-3-acetonitrile-4-methoxy-2-S-β-D-glucopyranoside (2) and N-methoxy-indole-3-acetonitrile-2-S-β-D-glucopyranoside (3), together with 11 known indole alkaloids were isolated from the roots of Isatis indigotica Fort. (Cruciferae). The structures of 1–3 were elucidated on the basis of mass spectrometry and extensive 1D and 2D NMR spectroscopy. All of the isolated compounds were tested for inhibitory activity against LPS-induced nitric oxide production in RAW 264.7 macrophages. A plausible biosynthesis pathway of 1–3 is also proposed.     

Effects of Morus alba leaf extract on the production of nitric oxide, prostaglandin E2 and cytokines in RAW264.7 macrophages

Morus alba leaf methanolic extract and its fractions (chloroform, butanol, and aqueous fractions) were found to inhibit NO production in LPS-activated RAW264.7 macrophages without an appreciable cytotoxic effect at concentration from 4 to 100 microg/ml. LPS-induced PGE2 production was significantly reduced only by butanol fraction. In addition, M. alba leaf extract and its fractions significantly decreased the production of TNF-alpha. These findings suggest that M. alba leaf extract seems to be able in suppressing inflammatory mediators. Moreover, the inhibitory activities on COX-2 and iNOS of its butanol fraction are warranted for further elucidation of active principles for development of new antiinflammatory agents.     

3, 4-seco-Labdane diterpenoids from the leaves of Callicarpa nudiflora and their inhibitory effects on nitric oxide production

As a part of our ongoing search for bioactive compounds from the leaves of Callicarpa nudiflora that inhibit nitric oxide (NO) production, the 90% EtOH fraction eluted by macroporous resin adsorption was found to show significant inhibitory activity against the production of NO in RAW 264.7 stimulated by lipopolysaccharide (LPS). Bioactivity-guided isolation of the fraction yielded three new bioactive diterpenoids, named ent-3, 4-seco-14-carbonyl-15, 16-epoxy-4(18), 8(17), 13(14)-labdatrien-3-oic acid (1), syn-3, 4-seco-12R-hydroxy-15, 16-epoxy-4(18), 8(17), 13(16), 14(15)-labdatetraen-3-oic acid (2) and syn-3, 4-seco-12S-hydroxy-15, 16-epoxy-4(18), 8(17), 13(16), 14(15)-labdatetraen-3-oic acid (3). Their structures and configurations were elucidated by comprehensive spectroscopic analysis. All the three compounds exhibited potent inhibitory activity on NO production in LPS-activated RAW 264.7 macrophage cells.     

Sesquiterpenes from Ulmus davidiana var. japonica with the inhibitory effects on lipopolysaccharide-induced nitric oxide production

Investigation of antiinflammatory constituents of the stem and root barks of Ulmus davidiana var. japonica resulted in the isolation of three guaiane type sesquiterpenes, torilin, 1-hydroxytorilin, together with a new derivative, (1beta, 7beta, 8beta, 10beta)-1,8,11-trihydroxy-4-guaien-8-angeloyl-3-one named 1-hydroxytorilin A. All the three sesquiterpenes inhibited lipopolysaccharide-induced nitric oxide production in murine microglial BV2 cells.     

Chemical constituents of Morus alba L. and their inhibitory effect on 3T3-L1 preadipocyte proliferation and differentiation

Mulberry leaf, an important traditional Chinese medicine, possesses many biological activities, including effects of anti-obesity. However, which constituents of mulberry leaf are responsible for its anti-adipogenic action is unclear. This study primarily investigated the chemical constituents from mulberry leaf and their bioactivity on the proliferation and differentiation of 3T3-L1 preadipocytes. A new flavane derivative, (2S)-4′-hydroxy-7-methoxy-8-prenylflavan (1), together with twelve known compounds including three flavanes (2–4), three chalcones (5–7), two flavones (8–9), two benzofurans (10–11) and two coumarin (12–13) was isolated from mulberry leaf. The structure of the new compound was elucidated by various spectroscopic methods including UV, HR-ESI-MS, ¹H and ¹³C NMR and CD. The results of activity screening showed that compound 2, 6 and 7 inhibited the proliferation and differentiation of 3T3-L1 preadipocytes.     

Antimicrobial activity of 2-arylbenzofurans from Morus species against methicillin-resistant Staphylococcus aureus

Nine 2-arylbenzofurans isolated from Morus species were tested for their antimicrobial activities against methicillin-sensitive Staphylococcus aureus (MSSA), methicillin-resistant S. aureus (MRSA), Micrococcus luteus, Bacillus subtilis, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa. Among these compounds, chalcomoracin (a leaf phytoalexine of mulberry tree) exhibited considerable antibacterial activity against MRSAs (MICs 0.78 μg/ml).     

Inhibition of nitric oxide production on LPS-activated macrophages by kazinol B from Broussonetia kazinoki

The ethyl acetate soluble fraction of Broussonetia kazinoki at 50 microg/ml showed a significant inhibition (78.2%) of nitric oxide (NO) in lipopolysaccharide activated macrophages. Kazinol B, an isoprenylated flavan, was identified as the active principle. It inhibits the NO synthesis with an IC(50) of 21.6 microM.     

桑树查尔酮异构酶基因的克隆与原核表达分析

以果叶兼用新桑品种‘嘉陵30号’的果实为材料,采用同源克隆法和抑制PCR法克隆桑树查尔酮异构酶基因(MaCHI)全序列.该基因的基因组序列全长2 402 bp,包含2 112 bp长的ORF和290 bp长的3'UTR序列,ORF由4个外显子和3个内含子组成,该基因编码219个氨基酸.预测MaCHI编码蛋白质的分子质量为23.8 ku,理论等电点为5.29.采用RT-PCR法分析MaCHI在不同组织中的表达水平,在叶片和果中的表达量较高,在根中表达量较低.构建pET-28a(+)-MaCHI原核表达重组质粒,并在大肠杆菌中诱导表达.     

Sesquiterpenes from Vladimiria souliei and their inhibitory effects on NO production

As a part of our ongoing search for plant-derived compounds that inhibit nitric oxide production, the methanol extract of the roots of Vladimiria souliei was found to show significant inhibitory effects on INF-γ-induced nitric oxide production in murine macrophage RAW264.7 cells. Bioactivity-guided isolation of the extract yielded two most active sesquiterpenes, including a new compound, named souliene A (1) and alismol (2). Their structures were elucidated by spectroscopic methods (IR, ESIMS, HRESIMS, 1D and 2D NMR). Two isolates showed promising inhibitory effects on INF-γ-induced nitric oxide production in murine macrophage RAW264.7 cells.     

Screening of Indonesian medicinal plants for inhibitor activity on nitric oxide production of RAW264.7 cells and antioxidant activity

Traditional Indonesian medicinal plants were screened for their inhibitory effects on the nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages and for the antioxidant activity through the evaluation of free radical scavenging effect and reducing power. The results of screening indicated that 50 methanolic extracts inhibited (>50%) lipopolysaccharides (LPS)-induced NO release from RAW264.7 cells at 50 microg/ml, with 18 having greater than 100% inhibition. At 200 microg/ml, 61 methanol extracts exhibited inhibitory activity (>50%), with 45 showing greater than 100% inhibition. In addition, the free radical scavenging effects of 6 methanolic extracts were found to be more than 50% for extract concentration of 0.5 mug/ml. The results indicate that the extracts contain active compounds that inhibit NO release and scavenge free radical.     

Variations in amounts of carbohydrates, amino acids and adenine nucleotides in mulberry tree (Morus alba L.) stems during transitional phases of growth

Quantitative changes in carbohydrates, amino acids and adenine nucleotides in the stems of mulberry trees (Morus alba L., cv. Shin-ichinose) were followed from spring to early summer and from autumn to early spring. Both ATP and ADP content of stems increased before bud break, whereas the content of sucrose, the most abundant sugar among the stem carbohydrates, decreased. The sucrose content fell to its lowest value at the beginning of May, and then increased rapidly, whereas the starch content decreased suggesting consumption of reserve carbohydrate and simultaneous accumulation of current photosynthate. This was confirmed by studies in which reserve carbohydrates were labeled with (14)CO(2). Proline content of stems increased from the time of leaf shedding until early spring. Although it was the most abundant amino acid at the time of bud break, proline rapidly decreased as new shoots developed and was hardly detectable by the beginning of May. The asparagine and arginine contents increased transiently following bud break, and then decreased toward summer. Transient increases in glutamine and arginine were noted at the time of leaf shedding.     

Cycloartane triterpenoids from the stems of Schisandra glaucescens and their bioactivity

Five new cycloartane triterpenoids, schiglausins K-O (1-5), including one hexanortriterpenoid (1) and one octanortriterpenoid (2), as well as two known compounds (6-7), were isolated from the stems of Schisandra glaucescens Diels. Their structures were elucidated on the basis of spectroscopic methods, including extensive NMR spectra. Compounds 2-7 were tested for their FXR agonistic and antagonistic effects. Compound 7 exhibited significant antagonistic effect against FXR with IC(50) of 1.50 μM.     

Lathyrane diterpenes from Euphorbia prolifera and their inhibitory activities on LPS-induced NO production

A new lathyrane diterpene (1), an unreported spectroscopic data lathyrane diterpenene (2), and two known analaogues (3 and 4) have been isolated from Euphorbia prolifera. Their structures were elucidated as (12E,2S,3S,4R,5R,6S,9S,11S,15R)-3-butyryloxy-5,15-diacetoxy-6,17-epoxylathyra- 12-en-14-one (1), (12E,2S,3S,4R,5R,6S,9S,11S,15R)-3-propionyloxy-5,15-diacetoxy-6,17- epoxylathyra-12-en-14-one (2), (12E,2S,3S,4R,5R,6S,9S,11S,15R)-3-benzoyloxy-5,15-diacetoxy -6,17-epoxylathyra-12-en-14-one (3), and 15-O-acetyl-17-hydroxyjolkinol (4) by spectroscopic methods (IR, ESIMS, HR-ESIMS, NMR, and X-ray crystallography). The inhibitory activities on LPS-induced NO production of these diterpenes were evaluated and compounds 1, 3 and 4 showed inhibitory effects.     

Phenolic glycosides from Dodecadenia grandiflora and their glucose-6-phosphatase inhibitory activity

Phytochemical investigation of Dodecadenia grandiflora leaves led to the isolation and identification of three phenolic glycosides, designated 1-[(4′-O-(E)-p-coumaroyl)-β-d-glucopyranosyl]-oxy-2-phenol (1), 1-[(6′-O-(E)-p-coumaroyl)-β-d-glucopyranosyl]-oxy-2-phenol (2) and 1-[O-β-d-glucopyranosyl(1→2)-β-d-glucopyranosyl]-oxy-2-phenol (3), along with nine known compounds. Compounds 1, 2, 5 and 9 exhibited significant glucose-6-phosphatase inhibitory activity (63.7, 66.9, 82.9 and 85.4%) with IC₅₀ values of 88.5, 81.0, 51 and 50 μM respectively. On the basis of biological results, a structure–activity relationship has been discussed.     

Excretion of tectoridin metabolites in rat urine and bile orally administrated at different dosages and their inhibitory activity against aldose reductase

This study investigated the urinary and biliary excretion of tectoridin, a major active isoflavonoid found in the flowers of Pueraria thomsonii Benth. and the rhizomes of Belamcanda chinensis (L.) DC. Using UHPLC/Q-TOFMS, seven glucuronides and/or sulfated metabolites and four Phase I metabolites were simultaneously quantified in rat urine after oral administration of tectoridin at 100 and 200 mg/kg. Over a 72-h period, 14.2% and 14.7% of the tectoridin were excreted as eleven metabolites in urine, among which, two major metabolites tectorigenin-7-O-β-D-glucuronide (Te-7G) and tectorigenin accounted for 5.5–5.5% and 4.3–4.4%. Furthermore, the cumulative excretion of four glucuronides and sulfated metabolites in bile accounted for 7.3% and 3.9% of the dose within 60 h, among which, Te-7G and tectorigenin-7-O-glucuronide-4′-O-sulfate (Te-7G-4′S) accounted for 2.3–3.0% and 1.4–3.9%, respectively. The results indicate that the urine was the primary elimination route, and glucuronidation after deglycosylation at C-7 position was the major metabolic pathway of tectoridin in vivo. Moreover, the inhibitory activities of tectoridin and its five metabolites on rat lens aldose reductase were confirmed (IC₅₀: 1.4–15.5 μM), whereas irisolidone-7-O-glucuronide (Ir-7G) and irisolidone showed little activity.     

neo-Clerodane diterpenes from Ajuga decumbens and their inhibitory activities on LPS-induced NO production

A phytochemical investigation of the whole plants of Ajuga decumbens led to the isolation of three new (1, 2a, and 2b) and three known (3a−3c) neo-clerodane diterpenes. Their structures were elucidated by spectroscopic data analysis (IR, ESIMS, HR-ESIMS, 1D and 2D NMR), and the structure of 1 was confirmed by X-ray crystallography. The inhibitory activities on LPS-induced NO production of the six diterpenes were evaluated and compounds 2a, 2b, and 3a showed inhibitory effects.     

Polar cardenolide monoglycosides from stems and twigs of Nerium oleander and their biological activities

Twelve polar cardenolide monoglycosides, 1, 2, 4?C13, and oleagenin (3) were isolated from the methanol extract of stems and twigs of Nerium oleander. Among these, oleagenin (3) and cardenolide monoglycosides named cardenolide B-1 (1) and cardenolide B-2 (2) were isolated from natural sources for the first time. The in vitro antiinflammatory activity of compounds 1?C13 was examined on the basis of inhibitory activity against the induction of the intercellular adhesion molecule-1 (ICAM-1). Compounds 4?C7 were active at an IC₅₀ value of less than 0.4 ??M. The cytotoxic activity of compounds 1?C13 was evaluated against three human cell lines: normal human fibroblast cells (WI-38), malignant tumor cells derived from WI-38 (VA-13), and human liver tumor cells (HepG2). Compounds 4, 6, and 7 were active toward these three cell lines at IC₅₀ values of less than 0.7 ??M, and compounds 5 and 8 were active toward the cell lines at IC₅₀ values of less than 1.5 ??M. The multidrug resistance (MDR) cancer-reversal activity of compounds 1?C13 was evaluated on the basis of the amount of calcein accumulated in MDR human ovarian cancer 2780AD cells in the presence of each compound. Compound 1 and 12 showed significant effects on calcein accumulation.     

Five novel naphthylisoquinoline alkaloids with growth inhibitory activities against human leukemia cells HL-60, K562 and U937 from stems and leaves of Ancistrocladus tectorius

Two new 7,6′-coupled naphthylisoquinolines, namely ancistrotectorines A (1) and B (2), two new 5,3′-coupled naphthylisoquinolines, namely ancistrotectorines C (3) and D (4), and one new 7,8-coupled naphthylisoquinoline, namely ancistrotectorine E (5), together with 9 known naphthylisoquinoline alkaloids, hamatine (6), ancistrobertsonine B (7), ancistrocladinine (8), hamatinine (9), ancistrotanzanine A (10), ancistrotanzanine B (11), ancistrotectoriline B (12), 7-epi-ancistrobrevine D (13), and ancistrotectorine (14), were isolated from the 70% EtOH extract of Ancistrocladus tectorius. Their structures were elucidated based on the extensive analysis of spectroscopic data (1D, 2D NMR and MS). Compound 5 exhibited inhibitory activities against HL-60, K562 and U937 cell lines with IC₅₀ values of 1.70, 4.18 and 2.56 μM respectively.