Sephadex LH-20 Research Articles

Molecular docking, molecular dynamics, MM/PBSA approaches and bioactivity studies of nepetanudoside B isolated from endemic Nepeta aristata

Nepetanudoside B (NNB) was isolated from aerial parts of endemic Nepeta aristata crude extract (CH3OH-CHCl3) using silica gel (n-hexane, methanol, ethyl acetate, and dichlorometane, respectively) and sephadex LH-20 (65% Methanol-35% Chloroform) column chromatographies. Preparative-HPLC was used to purify NNB after activity-guided isolation of methanol sub-fractions with enzyme inhibitory and DNA protective properties. The NNB was determined using 1H,13C, COSY, HSQC, HMBC, and LC-MS/MS. The study compared the effects of NNB with conventional drugs in terms of its ability to inhibit enzymes such as urease, α-amylase, carbonic anhydrase (CA), lipase, α-glucosidase, and tyrosinase, as well as its ability to protect DNA. Enzyme kinetic and molecular docking were also used to evaluate this. NNB exhibited the best inhibitory activity on urease (1.28 ± 0.00 µg/mL), lipase (5.83 ± 0.10 µg/mL), BChE (3.73 ± 0.46 µg/mL), tyrosinase (7.39 ± 0.00 µg/mL), α-glucosidase (10.95 ± 0.00 µg/mL), α-amylase (22.11 ± 1.03 µg/mL) and AChE (25.68 ± 3.32 µg/mL), respectively. NNB has higher MolDock scores with binding energy in α-glucosidase (-233) and BChE (-8.90 kcal/mol). In enzyme kinetics studies, it was determined that urease, AChE, α-glucosidase, lipase, and CA were non-competitive , while BChE and tyrosinase were competitive inhibition mechanisms. Their Ki values were calculated as 0.09, 0.24, 0.09, 0.10, 0.08, 0.05, and 0.07 mM, respectively. Molecular dynamics simulation studies were performed for the interactions of NNB-BChE with MM/PBSA binding free energey RMSD, RMSF, Rg, SASA, and also the number of hydrogen bonds was calculated. The suitability and effectiveness of NNB have been proven in the food and pharmaceutical industries. The NNB molecule may lead to development studies as a BChE inhibitor. Communicated by Ramaswamy H. Sarma

New Polyene Macrolide Compounds from Mangrove-Derived Strain Streptomyces hiroshimensis GXIMD 06359: Isolation, Antifungal Activity, and Mechanism against Talaromyces marneffei.

Mangrove-derived actinomycetes represent a rich source of novel bioactive natural products in drug discovery. In this study, four new polyene macrolide antibiotics antifungalmycin B-E (1-4), along with seven known analogs (5-11), were isolated from the fermentation broth of the mangrove strain Streptomyces hiroshimensis GXIMD 06359. All compounds from this strain were purified using semi-preparative HPLC and Sephadex LH-20 gel filtration while following an antifungal activity-guided fractionation. Their structures were elucidated through spectroscopic techniques including UV, HR-ESI-MS, and NMR. These compounds exhibited broad-spectrum antifungal activity against Talaromyces marneffei with minimum inhibitory concentration (MIC) values being in the range of 2-128 μg/mL except compound 2. This is the first report of polyene derivatives produced by S. hiroshimensis as bioactive compounds against T. marneffei. In vitro studies showed that compound 1 exerted a significantly stronger antifungal activity against T. marneffei than other new compounds, and the antifungal mechanism of compound 1 may be related to the disrupted cell membrane, which causes mitochondrial dysfunction, resulting in leakage of intracellular biological components, and subsequently, cell death. Taken together, this study provides a basis for compound 1 preventing and controlling talaromycosis.

Isolation, identification and α-glucosidase inhibition evaluation of a phenolic compound from Elaeocarpus mastersii King leaves

Elaeocarpus mastersii King is an Indonesian folk medicine with high phenolic content. In this study, the E. mastersii King leaves were extracted using methanol solvent through maceration. The methanol extract was continuously fractionated by the liquid-liquid extraction using hexane, dichloromethane (DCM), and ethyl acetate solvents, respectively. The DCM fraction was purified by column chromatography using Silica gel 60 and Sephadex LH-20 as the stationary phase and monitored by thin-layer chromatography. This isolation process led to obtaining a pure compound with the appearance of white crystalline powder and a melting point of 198-199℃. This compound absorbed the UV light at the wavelength of 229 and 274 nm, as well as produced the bathochromic shift by the addition of NaOH reagent indicating the aromatic compound with the hydroxyl (OH) substituent. The IR spectrum confirmed the presence of aromatic and OH groups along with other functional groups, such as carbonyl (vmax 1693 cm-1) and alkane (C-H, vmax 2953 cm-1). The NMR data suggested the isolated compound as methyl gallate. Furthermore, the m/z value of the isolated compound was 185.04416 Da [M+H]+. It confirmed the monoisotopic mass of the methyl gallate (184.0362 Da). However, biological evaluation as an α-glucosidase inhibitor showed no positive activity.

Triterpenoids in Sorbus pohuashanensis suspension cell treated with yeast extract

This study induced biological stress in Sorbus pohuashanensis suspension cell(SPSC) with yeast extract(YE) as a bio-tic elicitor and isolated and identified secondary metabolites of triterpenoids produced under stress conditions. Twenty-six triterpenoids, including fifteen ursane-type triterpenoids(1-15), two 18,19-seco-ursane-type triterpenoids(16-17), four lupine-type triterpenoids(18-21), two cycloartane-type triterpenoids(22-23), and three squalene-type triterpenoids(24-26), were isolated and purified from the methanol extract of SPSC by chromatography on silica gel, MCI, Sephadex LH-20, and MPLC. Their structures were elucidated by spectroscopic analyses. All triterpenoids were isolated from SPSC for the first time and 22-O-acetyltripterygic acid A(1) was identified as a new compound. Selected compounds were evaluated for antifungal, antitumor, and anti-inflammatory activities, and compound 1 showed an inhibitory effect on NO production in LPS-induced RAW264.7 cells.

Color characteristics and pyrolysis volatile properties of main colored fractions from the Maillard reaction models of glucose with three amino acids

The effects of hydrosoluble colored fractions generated in the Maillard reaction models on the volatiles of pyrolytic Maillard reaction products was investigated. The hydrosoluble colored fractions obtained by reaction of glucose with glycine (Gly-Glc), proline (Pro-Glc) and phenylalanine (Phe-Glc) respectively at 125 °C were separated using Sephadex LH-20 Gel column. The colored fraction with large molecular weight had a higher contribution to the overall color and more glucose residue structures in main skeleton than that with small molecular weight in the same model. Gel permeation chromatography analysis indicated that these colored fractions had continuous relative molecular weight distribution (1000–80000 Da). Pyrolysis - gas chromatography - mass spectrometry determination (pyrolysis temperature: 160∼900 °C) suggested that furans and pyrroles were the main volatiles of two Gly-Glc fractions after pyrolysis, and the key pyrolysis volatiles of Pro-Glc fraction were furans, pyrroles, pyran, indole and cyclopentenes, which gave a freshly baked bread, nutty, coffee-like and sweet caramel-like odor. Especially phenyl-containing compounds seemed typical indicators of Phe-Glc fraction pyrolysis products, exhibiting a benzene-like, tallowy, sweet, balsamic and floral odor. The hydrosoluble colored fractions produced by different Maillard reactions had different pyrolytic volatile properties and were suitable for the development of bakery food improver with different color and flavor requirements.

Bioassay-guided discovery and identification of new potent α-glucosidase inhibitors from Morus alba L. and the interaction mechanism

Ethnopharmacological relevanceMorus alba L. (mulberry) is a well-known medicinal species that has been used by herbalist doctors for the treatment of diabetes for a long history, and modern ethnopharmacological studies have demonstrated the ameliorating effects of different mulberry extracts toward diabetes-related symptoms and identified a number of α-glucosidase inhibitors as hypoglycemic ingredients. Aim of the studyThe present study aims to explore new potent α-glucosidase inhibitors from the root bark of M. alba (known as Sang-Bai-Pi in traditional medicine) based on an in vivo antidiabetic evaluation of its extract fractions and further characterize the preliminary mechanism of the new active constituents. Materials and methodsα-Glucosidase inhibitory assay and diabetic mice model were used to locate and evaluate the active fractions from the extract. Diverse separation techniques (e.g. Sephadex LH-20 column chromatograph (CC) and HPLC) and spectroscopic methods (e.g. MS, NMR and ECD) were employed to isolate and structurally characterize the obtained constituents, respectively. Fluorescence quenching, kinetics and molecular docking experiments were conducted to investigate the enzyme inhibitory mechanism of the active compounds. ResultsThe 80% ethanol eluate from the macroporous resin CC exerted good antidiabetic effects in the tested mice. Fifty-two flavonoids including 22 new ones were then separated and identified, and most of them showed strong inhibition against α-glucosidase with their structure-activity relationship being also discussed. The four new most active ingredients were further characterized to be mixed type of α-glucosidase inhibitors, and their binding modes with the enzyme were also explored. ConclusionsOur current work has demonstrated that the root bark of M. alba is an extremely rich source of flavonoids as potent α-glucosidase inhibitors and potential antidiabetic agents, which makes it a promising candidate species to develop new natural remedies for the prevention and treatment of diabetes.

Limonoids from the Branches and Leaves of Aglaia lawii and Their Cytotoxic Activities.

Three undescribed limonoids (1-3), named aglaians G-I, and one new natural product azedaralide (4), together with nine known analogues (5-13) were isolated from the branches and leaves of Aglaia lawii by RP C18 column, silica gel column, Sephadex LH-20 column chromatography and preparative HPLC. The structures of the new compounds were elucidated by IR, HRESIMS, 1D, 2D NMR, electronic circular dichroism (ECD) calculations and X-ray crystallography diffraction analysis. The results of bioassay showed that the compound 12 exhibited potential inhibitory activity against six human tumor cell lines (MDA-MB-231, MCF-7, Ln-cap, A549, HeLa and HepG-2) with IC50 values as 8.0-18.6 μM.

CHEMICAL STRUCTURE OF THE POLYPHENOL COMPOSITION IN THE LEAVES OF PONTAIN HAWTHORN CRATAEGUS PONTICA

The chemical composition (polyphenols) of the leaves of the Crataegus pontica K. Koch plant has been studied. The process of isolating polyphenols from plant raw materials includes several stages: extraction of raw materials, processing of the extract with organic solvents, evaporation, precipitation of the sum of polyphenols, purification. The increase in the efficiency of the use of raw materials is achieved mainly at the first stage – extraction. The yield of the sum of polyphenols was studied depending on: the composition of the extractant, the extraction modulus, the multiplicity of extraction, the ratio of raw materials – extractant, temperature, thickening conditions, treatment of the aqueous residue with organic solvents, the conditions of precipitation of the sum of polyphenols and their drying. As a result, optimal conditions for the isolation of polyphenols from plant raw materials were selected, and the found condition obtained the sum of polyphenols from the autumn leaves of pontic hawthorn with a yield of 4.5%. Chromatographic study of the isolated fractions revealed that the polyphenols of the ethyl acetate fraction are mainly represented by monomeric catechins, flavonolams and tannins. The sum of polyphenols from the ethyl acetate fraction was divided on a column with sephadex LH-20 using chloroform-methanol as an eluent in various ratios (10 : 1→3 : 1). The structures of the isolated compounds were determined using physicochemical methods. As a result, more than 12 polyphenols were isolated from the leaves of pontic hawthorn, such as quercetin-3-O-β-D-galactopyranoside (hyperoside), quercetin-3-rutinoside, 3,5,7,3',4'-pentaoxyflavone (quercetin), 2-(3,4-dimethoxyphenyl)-7methoxy-3,4-dihydro-2H-chromene-3,4,5,6-tetrol, aрigenin-6-C-glycoside, procyanidin B, (+)-catechin (5,7,3',4'-tetraoxyflavan-3-ol), (-)-epicatechin (5,7,3',4'-tetraoxyflavan-3-ol), ellagic acid, gallic acid, 1-O-galloyl-4-O-catechin-β-D-glucose and caffeic acid ester with 2,3-dihydroxyglutaric acid. Of these, the last two substances were first isolated from this plant.

Chemical constituents from Salacia polysperma

Nine compounds were isolated from the 90% ethanol extract of Salacia polysperma by silica gel, Sephadex LH-20 column chromatography, together with preparative HPLC methods. Based on HR-ESI-MS, MS, 1D and 2D NMR spectral analyses, the structures of the nine compounds were identified as 28-hydroxy wilforlide B(1), wilforlide A(2), 1β,3β-dihydroxyurs-9(11),12-diene(3),(-)-epicatechin(4),(+)-catechin(5),(-)-4'-O-methyl-ent-galloepicatechin(6), 3-hydroxy-1-(4-hydroxy-3-methoxy-phenyl)propan-1-one(7),(-)-(7S,8R)-4-hydroxy-3,3',5'-trimethoxy-8',9'-dinor-8,4'-oxyneoligna-7,9-diol-7'-aldehyde(8), and vanillic acid(9). Compound 1 is a new oleanane-type triterpene lactone. Compounds 1, 3, 4, 7-9 were isolated from the Salacia genus for the first time. All compounds were assayed for their α-glucosidase inhibitory activity. The results suggested that compound 8 exhibited moderate α-glucosidase inhibitory activity, with an IC_(50) value of 37.2 μmol·L~(-1), and the other compounds showed no α-glucosidase inhibitory activity.

Chemical constituents of Helleborus thibetanus

The chemical constituents of Helleborus thibetanus were isolated and purified by silica gel column chromatography, Sephadex LH-20 gel column chromatography, and semi-preparative RP-HPLC, and the structures of all compounds were identified by modern spectrographic technology(MS, NMR). The MTT method was used to measure the cytotoxicity of compounds 1-8. Twelve compounds were isolated from the roots and rhizomes of H. thibetanus and were identified as(25R)-22β,25-expoxy-26-[(O-β-D-glucopyranosyl)oxy]-1β,3β-dihydroxyfurosta-5-en(1), β-sitosterol myristate(2), β-sitosterol lactate(3), β-sitosterol 3-O-β-D-glucopyrannoside(4), 4,6,8-trihydroxy-3,4-dihydronaphthalen-1(2H)-one(5), 1,3,5-trimethoxybenzene(6), 7,8-dimethylbenzo pteridine-2,4(1H,3H)-dione(7), 1H-indole-3-carboxylic acid(8), p-hydroxy cinnamic acid(9), lauric acid(10), n-butyl α-L-arabinofuranoside(11) and methyl-α-D-fructofuranoside(12), respectively. Among them, compound 1 is a new compound and named thibetanoside L; compounds 2, 5-8, 11 are first isolated from the family Ranunculaceae; compound 12 is isolated from the genus Helleborus for the first time. The results of MTT assay showed that the IC_(50) values of compounds 1-8 against HepG2 and HCT116 cells were greater than 100 μmol·L~(-1).

Anti-Inflammatory Ergosteroid Derivatives from the Coral-Associated Fungi Penicillium oxalicum HL-44

To obtain the optimal fermentation condition for more abundant secondary metabolites, Potato Dextrose Agar (PDA) medium was chosen for the scale-up fermentation of the fungus Penicillium oxalicum HL-44 associated with the soft coral Sinularia gaweli. The EtOAc extract of the fungi HL-44 was subjected to repeated column chromatography (CC) on silica gel and Sephadex LH-20 and semipreparative RP-HPLC to afford a new ergostane-type sterol ester (1) together with fifteen derivatives (2–16). Their structures were determined with spectroscopic analyses and comparisons with reported data. The anti-inflammatory activity of the tested isolates was assessed by evaluating the expression of pro-inflammatory factors Tnfα and Ifnb1 in Raw264.7 cells stimulated with LPS or DMXAA. Compounds 2, 9, and 14 exhibited significant inhibition of Ifnb1 expression, while compounds 2, 4, and 5 showed strong inhibition of Tnfα expression in LPS-stimulated cells. In DMXAA-stimulated cells, compounds 1, 5, and 7 effectively suppressed Ifnb1 expression, whereas compounds 7, 8, and 11 demonstrated the most potent inhibition of Tnfα expression. These findings suggest that the tested compounds may exert their anti-inflammatory effects by modulating the cGAS-STING pathway. This study provides valuable insight into the chemical diversity of ergosteroid derivatives and their potential as anti-inflammatory agents.

Echinodorus macrophyllus: Acute toxicological evaluation of hydroxycinnamoyl derivatives from SF1 subfractions

Ethnopharmacological relevanceEchinodorus macrophyllus (Kunth.) Micheli (Alismataceae), known as chapéu-de-couro in Brazil, is popularly used to treat inflammatory diseases. We have previously demonstrated a significant reduction in the acute inflammation for the aqueous extract of E. macrophyllus (AEEm) and its ethanolic fraction (Fr20) and described that hydroxycinnamoyl derivatives present in SF1 (Fr20 subfraction) showed higher anti-inflammatory properties by mechanisms that include a reduction of TNF-α, IL-1β, CKCL1/KC, LTB4, and PGE2 levels in exudate. Aim of the studyThis work describes the acute toxicological effect of SF1 subfraction on SW mice treated orally for five days in the air pouch model by evaluating the hematological and biochemical determinations on the blood samples; the relative organ weight and its histopathological analysis; the liver genotoxicity assessment and the activity of liver enzymes from xenobiotic metabolism. Materials and methodsFr20 was earlier fractionated on the Sephadex LH-20 column, yielding mainly four subfractions, including SF1. The SF1 toxicity was evaluated in mice challenged with carrageenan on the air pouch inflammation model and orally treated for five days. The body weight was monitored daily, and the organs were weighed after the euthanasia. Hematological and biochemical determinations were carried out using specific commercial kits and following the protocols provided by the manufacturers. The organs were fixed, sectioned, processed for hematoxylin and eosin staining, and analyzed by light microscopy. Genotoxicity assessment was performed by the alkaline single-cell gel electrophoresis. Livers were processed for ethoxyresorufin-O-deethylase (EROD) and Glutathione S-transferase (GST) assays. ResultsSF1 exhibited low toxicity, as no significant discrepancy was observed in the relative weight of the body organs of mice. Moreover, the daily treatment with SF1 did not alter the number and percentage of red blood cells or hemoglobin concentration in the blood. The treatment with SF1 did not affect the creatinine concentration, but the 25 mg/kg dose reduced the plasma urea level and uric acid, suggesting its use in treating acute renal failure. The parameters analyzed did not present biochemical alterations indicative of liver disease. Regarding serum triglyceride and cholesterol levels, a significant decrease was detected in both parameters in mice treated with SF1. In addition, the histopathological analysis showed that inflammatory focus in the livers seemed more relevant in the control groups than in those treated. There were no significant changes in the renal or splenic tissues of animals treated with SF1. Treatment with SF1 also does not have a genotoxic effect on liver cells. ConclusionTreatment with SF1 showed no toxicity in mice at doses equivalent to those recommended for humans, which provides evidence of the safety of the therapeutic use of this subfraction.

Identification of flavonoids in Anoectochilus roxburghii and in vitro inhibition of key enzymes relevant to metabolic syndrome

The traditional Chinese herb Anoectochilus roxburghii (A. roxburghii) is well-known for its medicinal value, but few studies have reported its ability to combat metabolic syndrome activities in vitro. In this study, the extraction of seven compounds from A. roxburghii using 70% ethanol, RP-C18, thin layer chromatography (TLC), Sephadex LH-20, high performance liquid chromatography (HPLC) and subsequent identification of their structures through nuclear magnetic resonance (NMR). Among these compounds, kaempferol-3-O-rutinoside and isorhamnetin-3-O-α-l-rhamnopyranoside-7-O-β-d-glucopyranoside were obtained from A. roxburghii for the first time. Results showed that those compounds inhibited key enzymes involved in metabolic syndrome in vitro. Molecular docking and binding free energy calculations were employed to elucidate the inhibition mechanism of these compounds, while SwissADME was used to assess their drug-like characteristics. Notably, 9-hydroxy-megastigma-4,7-dien-3-one-9-O-β-d-glucopyranoside has the potential as an α-amylase inhibitor for the first time. This research enriches the compound library of A. roxburghii and provides new ideas for medication research and the development of metabolic syndrome.

Exploring separation patterns and mechanisms of proanthocyanidins in grape seeds and pomace with diverse molecular weights, compositions, and structures

The function of proanthocyanidins (PAs) relies on their structure and requires high-purity PAs. Though Sephadex LH-20 gel permeation chromatography (GPC) is expected to separate PAs based on structure, its usage rules and mechanisms remain unclear. This study delves into the PAs separation patterns on Sephadex LH-20, first confirming the purification mechanisms of PAs with various mean degrees of polymerization (DP) using the adsorption kinetic model. The study found that an increase in the molecular weight or mean DP of PAs results in decreased polarity, reduced hydrogen bonding actions, and intensified hydrophobic effect, causing delayed extraction of PAs on Sephadex LH-20, with galloylated PA as an exception, which was extracted first despite its high DP. Additionally, the principles for separating specific composition, such as monomers, dimers, etc., were evaluated. The study sheds light on enhancing the purification efficiency of PAs, thus advancing the precise separation technology of diverse proanthocyanidins.

Characterisation of pomegranate juice polyphenols identifies oenothein B as an endothelium-dependent vasodilator and regulator of endothelial function that enhances actions of oligomeric procyanidins

Abstract Background Cardiovascular disease mortality is reduced in individuals with high daily consumption of dietary polyphenols. Optimal daily intakes, choice of polyphenol, and mechanism of action have yet to be fully defined. Pomegranate juice (PJ) reduces blood pressure and improves endothelial function. These effects are attributed to the high levels of polyphenols, primarily ellagitannins. Purpose The aims of these investigations were (1) to identify PJ polyphenol(s) that altered endothelial function and could potentially lower blood pressure, and (2) to compare these endothelial actions to oligomeric procyanidins (OPC) purified from apple polyphenol extracts. Methods PJ polyphenols were purified by Sephadex LH-20 chromatography and RP-HPLC, and identified by LC-MS/MS and NMR. Fractions were screened for biological activity using cultured bovine aortic endothelial cells by measuring the degree of inhibition of endothelin-1 synthesis. Endothelial actions were further assessed by measuring changes in intracellular Ca2+ using FLIPR Ca2+-6 assay, qRT-PCR (EDN1, eNOS and KLF2 mRNAs), and NO synthesis (nitrite). Results Six PJ polyphenols were isolated. These were identified as punicalin, four punicalagin isomers, and oenothein B (OTB, a cyclic dimeric ellagitannin; LC-MS [M-H]-= 1567.1). At concentrations ≤5 µg/ml only OTB inhibited ET-1 synthesis (IC50 1.5 µg/ml). The identity of OTB was confirmed by NMR, and by comparison with OTB purified from Epilobium species. Further characterisation of the actions of OTB on endothelial cells showed pre-incubation with 1.25, 2.5 and 5 µg/ml OTB increased binding of biotinylated-OPC by 1.7, 2.3 and 3.8 fold. Subsequent studies of the interactions between OTB and OPC showed synergistic effects for decreases in ET-1 synthesis, EDN1 mRNA, and increases in KLF2 mRNA, eNOS mRNA and NO release. The effect of combining OTB (1 µg/ml) and OPC (1 µg/ml) lasted for over 24 h (relative to control: ET-1 -57%, EDN1 mRNA -63%, eNOS mRNA +69%, KLF2 mRNA +356%), when individually these polyphenols no longer had any significant effect. Intracellular Ca2+ responses also showed synergistic interactions, however the profile of [Ca2+] changes for OTB and OPC alone differed, suggesting separate signalling pathways initiating these endothelial responses. Endothelium-dependent vasodilator effects of OTB were confirmed on isolated mouse aorta (EC50 0.27 µg/ml). Conclusions These effects of OTB may underlie previously reported decreases in blood pressure and improvements in endothelial function after pomegranate juice consumption. Detailed understanding of the mechanism of action of OTB, and its interactions with OPC could lead to new therapeutic modalities for treating endothelial dysfunction in a broad spectrum of people at risk of cardiovascular disease.

Isolation, Characterization, and Antiproliferative Activity of Polymethoxy Flavones from Mentha dumetorum

Background: Natural products have been used commonly in the pharmaceutical industry as well as traditional medicine due to their bioactive contents. Objective: The objective of this study is to isolate the polymethoxy flavones (PMFs) and evaluate the antiproliferative activity. Methods: The PMFs were extracted from Mentha dumetorum with boiling hot water and then partitioned with hexane and the extract was subjected to chromatographic techniques such as Sephadex LH-20, silica gel, and preparative TLC, and HPLC to isolate the compounds. The structures of isolated compounds were elucidated by spectroscopic techniques such as 1H-NMR, 13C-NMR, LC-TOF-MS, and UV-Vis. The antiproliferative effects of isolated compounds were evaluated using a real-time cell analyzer–single plate (RTCA-SP) instrument against HeLa and HT29 cell lines. Results: The isolated PMFs were identified as xanthomicrol (1), 5-hydroxy-6,7,8,4'-tetrametoxy flavanone (2), 5-desmetil sinensetin (3), 5-demetil nobiletin (4), gardenin B (5), 5-hydroxy-6,7,8,3',4'- penta methoxy flavanone (6) and 5-hydroxy-6,7,4'-trimethoxy flavone (7). Compounds 2 and 5 were found to be the most active against both cell lines. Conclusion: The isolated compounds as well as the plant extract of Mentha dumetorum could be promising agents for the drug development process, especially drugs for cancer treatment. Moreover, isolation methods were developed for the corresponding compounds.

Research on Secondary Metabolites From the Soft Coral-Associated Fungus Aspergillus sydowii SCSIO41203 and Their Bioactivities

Objective: As part of our ongoing search for bioactive secondary metabolites, the Beibu Gulf soft coral-derived fungus Aspergillus sydowii SCSIO41203 was investigated, which led to the isolation of 15 compounds. Methods: These compounds were purified by chromatographic methods, including normal phase silica gel column chromatography, octadecylsilyl column chromatography, Sephadex LH-20, and high-performance liquid chromatography. Their structures were elucidated by nuclear magnetic resonance, and mass spectrometry and compared their data with those related metabolites in the literature. Results: The structures of 1 to 15 were identified as expansol F (1), expansol C (2), expansol D (3), ( Z)-5-(hydroxymethyl)-2-(6′-methylhept-2′-en-2′-yl)phenol (4), ( E)-5-(hydroxymethyl)-2-(6′-methylhept-2′-en-2′-yl)phenol (5), sydonol (6), (-)-(7 R,10 S)-10-hydroxysydowic acid (7), (-)-(7 R,10 R)- iso-10-hydroxysydowic acid (8), sydowic acid (9), 1-hydroxyboivinianin A (10), (+)-sydonic acid (11), 3-hydroxydiorcinol (12), cordyol C (13), ( R)-mevalonolactone (14), and 4-methyl-5,6-dihydropyren-2-one (15). These isolated compounds were evaluated for antibacterial, cytotoxic, and enzyme-inhibitory activities. Among them, compounds 1 to 3 and 13 showed moderate acetylcholinesterase inhibitory activity, with IC50 values of 3.9, 33.8, 11.5, and 19.8 μM, respectively, with the positive control (tacrine) of 0.5 μM.

A new allo-aromadendrane sesquiterpene from Dendrobium nobile

To study the chemical constituents in the non-alkaloid part of stems of Dendrobium nobile. The macroporous adsorption resin, MCI, silica gel, RP-C_(18), and Sephadex LH-20 gel, preparative thin layer chromatography, and preparative high-performance liquid chromatography(HPLC) were used to isolate and purify the compounds. The structures of the compound were determined according to the spectra data, physicochemical properties, and relevant references. A total of 8 compounds were isolated from D. nobile, which were soltorvum F(1), p-hydroxyphenylpropionic acid(2), vanillic acid(3), p-hydroxybenzoic acid(4), N-trans-cinnamic acid acyl-p-hydroxybenzene ethylamine(5),(+)-(1R,2S,3R,4S,5R,6S,9R)-2,11,12-trihydroxypicrotoxane-3(15)-lactone(6), dendronobilin H(7), soltorvum E(8). Compound 1 was a novel compound, named as soltorvum F. Compound 8 was isolated from Dendrobium species for the first time.

Two new phenylpropanoids from wine-processed Corni Fructus

Six compounds were isolated from aqueous extract of wine-processed Corni Fructus through silica gel, ODS column chromatography, Sephadex LH-20 gel column chromatography, reverse phase preparative HPLC and other chromatographic separation technologies. Their structures were identified with multiple spectroscopical methods including HR-ESI-MS, UV, IR, NMR and ECD and so on. Their structures were established as pinoresinoside B(1), cornusgallicacid A(2),(+)-isolariciresinol-9'-O-β-glucopyranoside(3),(-)-isolariciresinol 3α-O-β-D-glucopyranoside(4),(7R,8S)-dihydrodehydrodiconiferyl alcohol 9-O-β-D-glucopyranoside(5), and(-)-seco isolariciresinol-9'-O-β-D-glucopyranoside(6). Among them, compounds 1 and 2 were two new compounds. The biological activity evaluation results showed that compounds 2 and 6 had strong DPPH free radical scavenging ability, with EC_(50) values of(4.18±1.96) and(21.45±1.19) μmol·L~(-1), respectively. Compounds 1 and 2 had protective effects on H_2O_2-induced oxidative damage in NRK-52E cells in a dose-dependent manner, and the cell survival rate of compound 2 at 100 μmol·L~(-1) was 96.09%±1.77%.

A new xanthone from hulls of Garcinia mangostana and its cytotoxic activity

Eight compounds were isolated from ethyl acetate fraction of 80% ethanol extract of the hulls of Garcinia mangostana by silica gel, Sephadex LH-20 column chromatography, as well as prep-HPLC methods. By HR-ESI-MS, MS, 1D and 2D NMR spectral analyses, the structures of the eight compounds were identified as 16-en mangostenone E(1), α-mangostin(2), 1,7-dihydroxy-2-(3-methy-lbut-2-enyl)-3-methoxyxanthone(3), cratoxyxanthone(4), 2,6-dimethoxy-para-benzoquinone(5), methyl orselinate(6), ficusol(7), and 4-(4-carboxy-2-methoxyphenoxy)-3,5-dimethoxybenzoic acid(8). Compound 1 was a new xanthone, and compound 4 was a xanthone dimer, compound 5 was a naphthoquinone. All compounds were isolated from this plant for the first time except compounds 2 and 3. Cytotoxic bioassay suggested that compounds 1, 2 and 4 possessed moderate cytotoxicity, suppressing HeLa cell line with IC_(50) va-lues of 24.3, 35.5 and 17.1 μmol·L~(-1), respectively. Compound 4 also could suppress K562 cells with an IC_(50) value of 39.8 μmol·L~(-1).