demethylwedelolactone and wedelolactone

Ultra-high-pressure extraction combined with high-speed counter-current chromatography was employed to extract and purify wedelolactone and isodemethylwedelolactone from Ecliptae Herba. The operating conditions of ultra-high-pressure extraction were optimized using an orthogonal experimental design. The optimal conditions were 80% aqueous methanol solvent, 200 MPa pressure, 3 min extraction time and 1:20 (g/mL) solid-liquid ratio for extraction of wedelolactone and isodemethylwedelolactone. After extraction by ultra-high pressure, the extraction solution was concentrated and subsequently extracted with ethyl acetate; a total of 2.1 g of crude sample was obtained from 100 g of Ecliptae Herba. A two-phase solvent system composed of petroleum ether-ethyl acetate-methanol-water (3:7:5:5, v/v) was used for high-speed counter-current chromatography separation, by which 23.5 mg wedelolactone, 6.8 mg isodemethylwedelolactone and 5.5 mg luteolin with purities >95% were purified from 300 mg crude sample in a one-step separation. This research demonstrated that ultra-high-pressure extraction combined with high-speed counter-current chromatography was an efficient technique for the extraction and purification of coumestans from plant material.

Topics: Asteraceae; Chemical Fractionation; Coumarins; Countercurrent Distribution; Plant Extracts; Pressure

The anticancer properties of demethylwedelolactone (DWEL) and wedelolactone (WEL), which are naturally occurring coumestans, have not been well characterized. In this study, we investigated the anti-invasive effects of synthetic WEL and DWEL on human MDA-MB-231 breast cancer cells. We found that WEL and DWEL inhibited the anchorage-independent growth and also suppressed cell motility and cell invasion of MDA-MB-231 cells. In addition, WEL and DWEL reduced the activity and expression of matrix metalloproteinases (MMPs) involved in blocking the IκB-α/NFκB and MEK/ERK signaling pathways in MDA-MB-231 cells. Furthermore, DWEL suppressed the metastasis and lung colonization of the tumor cells in the nude mice. Altogether, these data suggest that DWEL derivatives exert anti-invasive growth effect on breast cancer cells.

Topics: Animals; Antineoplastic Agents; Breast Neoplasms; Cell Movement; Cell Proliferation; Coumarins; Dose-Response Relationship, Drug; Drug Screening Assays, Antitumor; Humans; Lung Neoplasms; Mice; Mice, Nude; Molecular Structure; Neoplasm Invasiveness; Signal Transduction; Structure-Activity Relationship; Tumor Cells, Cultured

We genetically modified Eclipta alba using Agrobacterium rhizogenes LBA 9402, with the aim of producing secondary metabolites with pharmacological properties against phospholipase A(2) and the myotoxic activities of snake venom. Extracts from in natura aerial parts and roots, both native and genetically modified (in vitro), were prepared and analysed by high-performance liquid chromatography. In natura materials showed the coumestan wedelolactone at higher concentration in the aerial parts, while demethylwedelolactone appeared at higher concentration in roots. Among the modified roots, clone 19 showed higher concentrations of these coumestans. Our results show that the in natura extracts of plants collected from Botucatu and Ribeirão Preto were efficient in inhibiting snake venom phospholipase A(2) activity. Regarding in vitro material, the best effect against Crotalus durissus terrificus venom was that of clone 19. Clone 19 and isolated coumestans (wedelolactone and demethylwedelolactone) inhibited the myotoxic activity induced by basic phospholipases A(2) isolated from the venoms of Crotalus durissus terrificus (CB) and Bothrops jararacussu (BthTX-I and II). The search for antivenom is justified by the need of finding active principles that are more efficient in neutralizing snake venoms and also as an attempt to complement serum therapy.

Topics: Animals; Bothrops; Brazil; Coumarins; Crotalid Venoms; Crotalus; Eclipta; Male; Mice; Phospholipases A; Plant Components, Aerial; Plant Extracts; Plant Roots; Plants, Genetically Modified; Rhizobium

A reversed-phase high performance liquid chromatographic method (RP-HPLC) was developed for the simultaneous determination of wedelolactone and isodemethylwedelolactone in Herba Ecliptae from different sources. The separation was performed on a Kromasil C18 column (250 mm x 4.6 mm, 5 microm), with methanol-0.5% acetic acid (55 45, v/v) as mobile phase at a flow rate of 1.0 mL/min. The detection wavelength was set at 351 nm and the volume of injection was 20 microL. There were good linear relationships between the mass concentrations and the peak areas of isodemethylwedelolactone and wedelolactone in the ranges of 1.6 - 32.0 mg/L (r = 0.999 5) and 5.6 - 112.0 mg/L (r = 0.999 8), respectively. The recoveries were found to be in the range of 97.5% - 98.2% and 99.0% - 100.2%, respectively. The results of the experiments have demonstrated that the established method is rapid and simple with good accuracy and reproducibility for the simultaneous determination of wedelolactone and isodemethylwedelolactone. The method is suitable for the quality control of Herba Ecliptae from different sources.

Topics: Chromatography, High Pressure Liquid; Coumarins; Dendrobium; Isomerism; Molecular Structure

Wedelolactone (WL) and demethylwedelolactone (DWL) isolated from Eclipta alba were tested in the trypsin inhibition bioassay (in vitro). Both compounds showed potent activity. IC(50) values of WL and DWL were found to be 2.9 and 3.0 microg/mL respectively.

Topics: Animals; Chickens; Coumarins; Eclipta; Humans; Inhibitory Concentration 50; Ovum; Phytotherapy; Plant Extracts; Protease Inhibitors; Trypsin

To study the chemical constituents of Eclipta prostrata (L).. The constituents of E. prostrata were systematically separated with the Bohlmann method and percolation and hot extraction methods, and various chromatographies. The structures were elucidated by chemical and spectroscopic means.. Ten compounds were isolated from the Eclipta prostrata. Their structures were determined as wedelolactone (1), demethylwedelolactone (2), isodemethylwedelolactone (3), alpha-formylterthienyl (4), strychnolactone (5), beta-sitosterol (6), nonacosanol (7), stearic acid (8), lacceroic acid (9), 3,4-dihydoxy benzoic acid (10). Fourteen ocmpounds, including hydrocarbons and its esters were identified by GC-MS from the least polar fractions.. Compound 3 is a new coumestan named isodemethylwedelolactone. Compounds 2-10 and compounds characterized by GC-MS analysis were obtained for the first time from Eclipta prostrata.

Topics: Coumarins; Eclipta; Molecular Structure; Plants, Medicinal

Topics: Animals; Cells, Cultured; Chemical and Drug Induced Liver Injury; Coumarins; India; Medicine, Ayurvedic; Plants, Medicinal; Rats