harpagoside and harpagide

The endophytic fungi from root,main stem,branch and leaf of Scrophularia ningpoensis were isolated from Zhejiang,whether these strains could yield harpagide or harpagoside were tested by HPLC and LC-MS. According to the morphological characteristic and the similarity of the nucleotide sequence of internal transcribed spacer( ITS) between r DNAs,the strains producing harpagide or harpagoside were identified. The results showed that 210 strains were isolated from the samples,which were classified into 9 orders,13 families and 17 genera by morphological study. Harpagide was detected in endogenous fungi ZJ17 and harpagoside was detected in endogenous fungi ZJ25 by HPLC coupled with LC-MS. ZJ17 was identified as Alternaria alternate and ZJ25 was identified as A.gaisen by its morphology and authenticated by ITS( ITS4 and ITS5 regions and the intervening 5. 8 S rDNA region).

Topics: China; DNA, Fungal; DNA, Ribosomal Spacer; Endophytes; Fungi; Glycosides; Iridoid Glycosides; Pyrans; Scrophularia

Inflammation does not only lead to pain and functio laesa in the affected tissue but is also implicated in the onset and progression of cardiovascular diseases and cancer. Many medicinal plants show anti-inflammatory properties yet plant-constituents and their effect on molecular pathways involved in the attenuation of inflammation as well as cell migration are only poorly understood. Harpagophytum procumbens DC. ex MEISN. is a potent plant used as an immune modulator in traditional herbal medicine. Aim of this study was to investigate the influence of harpagoside and harpagide on TNFα-secretion in undifferentiated and differentiated THP-1 cells under inflammatory conditions as well as their implication in cellular migration into inflamed tissue. We found that both iridoids decrease TNFα-secretion in PMA-differentiated THP-1 cells whereas undifferentiated cells were poorly affected. Yet, in undifferentiated cells harpagoside and harpagide induced mRNA-expression of certain proteins involved in leukocyte transmigration. Especially TNFα and ICAM-1 mRNA-expression was positively affected after 3h and expression could be maintained on high levels even after 48h. L-selectin and PSGL-1 were strongly induced after 48h of stimulation. This ambiguous effect of harpagoside and harpagide highlights their immune modulatory function by facilitating cell migration into the inflamed tissue, whereby in consequence the anti-inflammatory activity of the resident macrophages was also found to be promoted.

Topics: Anti-Inflammatory Agents; Cell Differentiation; Cell Line; Cell Movement; Glycosides; Harpagophytum; Humans; Intercellular Adhesion Molecule-1; Interferon-gamma; Iridoid Glycosides; L-Selectin; Lipopolysaccharides; Macrophages; Membrane Glycoproteins; Monocytes; Plants, Medicinal; Pyrans; Tumor Necrosis Factor-alpha

An improved high-performance liquid chromatography with diode array detection combined with accelerated solvent extraction method was used to simultaneously determine six compounds in crude and processed Radix Scrophulariae samples. Accelerated solvent extraction parameters such as extraction solvent, temperature, number of cycles, and analysis procedure were systematically optimized. The results indicated that compared with crude Radix Scrophulariae samples, the processed samples had lower contents of harpagide and harpagoside but higher contents of catalpol, acteoside, angoroside C, and cinnamic acid. The established method was sufficiently rapid and reliable for the global quality evaluation of crude and processed herbal medicines.

Topics: Chromatography, High Pressure Liquid; Cinnamates; Coumaric Acids; Glucosides; Glycosides; Iridoid Glucosides; Iridoid Glycosides; Phenols; Pyrans; Quality Control; Scrophularia; Solvents; Trisaccharides

Harpagide (1) and harpagoside (2) are two iridoid glycosides existing in many medicinal plants. Although they are believed to be the main bioactive compounds related to the anti-inflammatory efficacy of these plants, the mechanisms of their anti-inflammatory activities remain unclear. The results of our present study showed that 1 and 2 had no effects on inhibitions of cyclooxygenase (COX)-1/2 enzyme activity, tumor necrosis factor-α (TNF-α) release, and nitric oxide (NO) production in vitro. However, the hydrolyzed products of 1 and 2 with β-glucosidase treatment showed a significant inhibitory effect on COX-2 activity at 2.5-100 μM in a concentration-dependent manner. Our further study revealed that the hydrolyzed 2 product was structurally the same as the hydrolyzed 1 product (H-harpagide (3)). The structure of 3 was 2-(formylmethyl)-2,3,5-trihydroxy-5-methylcyclopentane carbaldehyde, with a backbone similar to prostaglandins and COX-2 inhibitors such as celecoxib. All of them have a pentatomic ring with two adjacent side chains. The result of molecular modeling and docking study showed that 3 could bind to the COX-2 active domain well through hydrophobic and hydrogen-bonding interactions, whereas 1 and 2 could not, implying that the hydrolysis of the glycosidic bond of 1 and 2 is a pre-requisite step for their COX-2 inhibitory activity.

Topics: Animals; beta-Glucosidase; Cell Survival; Cyclooxygenase 1; Cyclooxygenase 2; Cyclooxygenase 2 Inhibitors; Glycosides; Humans; Hydrogen Bonding; Hydrolysis; Hydrophobic and Hydrophilic Interactions; Iridoid Glycosides; Macrophages; Mice; Models, Molecular; Molecular Structure; Osteoarthritis; Phytotherapy; Plant Preparations; Pyrans; Sodium Nitrite; Tumor Necrosis Factor-alpha

To develop an HPLC method for the simultaneous quantitation of five constituents in Scrophularia ningpoensis.. Samples were analyzed on an Agilent SB-C18 column(4.6 mm x 250 mm, 5 microm) eluted with acetonitrile and water containing 0.03% phosphate acid as mobile phases in a linear gradient mode. The flow rate was kept at 1.0 mL x min(-1), and the column temperature was set to 30 degrees C. The DAD detector wavelengths were 210, 280, 330 nm.. The linear ranges were 50-400 mg x L(-1) for harpagide, 1-40 mg x L(-1) for harpagoside, 1-20 mg x L(-1) for cinnamic acid, 0.5-4.5 mg x L(-1) for acteoside,1-60 mg x L(-1) for angoroside C, respectively. The average recoveries of the five constituents were 100.8% (RSD 0.62%), 101.7% (RSD 0.32%), 98.8% (RSD 0.48%), 99.9% (RSD 1.4%), 99.2% (RSD 1.1%), respectively.. Through the validation, the method was proved to be sensitive, accurate, repeatable, and can be used for quality control of the roots of S. ningpoensis.

Topics: Chromatography, High Pressure Liquid; Cinnamates; Coumaric Acids; Glucosides; Glycosides; Iridoid Glycosides; Phenols; Pyrans; Scrophularia; Trisaccharides

Potential interactions between herbal medicinal products and the cytochrome (CYP) P450 system are an important safety concern. We set out to develop a screening panel for assessing such interactions and use it to evaluate the interaction potential of devil's claw.. The panel consisted of luminescence-based inhibition assays for CYP1A2, 2C9, 2C19, 2D6 and 3A4, and a reporter gene (luciferase) assay for pregnane X receptor (PXR) activation and CYP3A4 induction. Caftaric acid and chlorogenic acid, two compounds with strong fluorescence quenching properties, were used to demonstrate the assay's resistance to interference. We tested 10 commercial devil's claw preparations as well as harpagoside and harpagide, two important constituents of devil's claw.. Five preparations were found to weakly inhibit CYP3A4 (IC50 124.2-327.6 µg/ml) and five were found to weakly activate PXR (EC50 10.21-169.3 µg/ml). Harpagoside and harpagide did not inhibit CYP3A4. In agreement with published data, bergamottin, a natural product known to interact with CYP3A4, was shown to inhibit CYP3A4 with an IC50 of 13.63 µm and activate PXR with an EC50 of 6.7 µm.. Devil's claw preparations are unlikely to have a clinically relevant effect on CYP function. The assay panel proved effective in screening devil's claw preparations, demonstrating its suitability for use with plant extracts. It showed superior sensitivity and resistance to interference.

Topics: Animals; Cattle; Cell Survival; Chromatography, High Pressure Liquid; Cytochrome P-450 Enzyme Inhibitors; Cytochrome P-450 Enzyme System; Drug-Related Side Effects and Adverse Reactions; Glycosides; Harpagophytum; Herb-Drug Interactions; Iridoid Glycosides; Luminescent Measurements; Phytotherapy; Plant Preparations; Plants, Medicinal; Pregnane X Receptor; Pyrans; Receptors, Steroid; Retinoid X Receptors

To develop a method for the determination of harpagide and harpagoside in Scrophulariae Radix (Xuanshen) by HPLC-UV under double wavelength, and to study the changes of these two constituents during processing, and to set the limitation of harpagide and harpagoside contents in crude drug and sliced pieces of Xuanshen.. The analyses were performed on an Agilent Technologies ZORBAX SB-C18 (4.6 mm x 250 mm, 5 microm) eluted with acetonitrile-water (containing 0.03% phosphoric acid) in gradient model. The flow rate was 1.0 mL x min(-1) . The column temperature was 25 degrees C. The UV detector wavelength was set at 210 nm before 13 min and then changed to 280 nm.. Harpagide and harpagoside were separated well. The linear calibration curves were obtained over of 0.0549 - 1.46 microg for harpagide (r = 0.9999, n =7) ,0.0225 - 0.900 microg for harpagoside (r = 0.9998, n = 9). The recoveries ( +/- RSD)% were 98.1 (+/- 2.4)% for harpagide and 98.8 (+/- 4.3)% for harpagoside. The contents of harpagide were 0. 277% - 0.620%, harpagoside were 0.078% - 0.362% in Xuanshen, and harpagide were 0.276% - 1.059%, harpagoside were 0. 059% - 0.183% in sliced Xuanshen, respectively. After the processing of Scrophulariae Radix, the content of harpagide increases 13.7% - 96.0%, while harpagoside decreases 11.0%-73.9%.. This method is simple, accurate, and can be used for the quality control of Scrophulariae Radix. We propose that the total content of harpagide and harpagoside in either crude drug or sliced pieces of Scrophulariae Radix should not be less than 0.45%.

Topics: Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Glycosides; Iridoid Glycosides; Magnoliopsida; Plant Roots; Pyrans; Spectrophotometry, Ultraviolet

The current work compared the relative efficacies of six commercial formulations of H. procumbens. Each formulation was assayed for the content of harpagoside (1), harpagide (2), verbascoside (3) and 8-O-p-coumaroyl harpagide (4) and, based on the recommended dosages, the total daily amounts were determined and used to establish anti-/proinflammatory (A/P) factors. The formulations were compared using ex vivo porcine skin for their activities towards COX-2 by Western blotting. The results showed great variation in the amounts of compounds 1-4 within the six formulations examined. The relative proportions of 1-4 also varied widely between the products and this inconsistency was reflected in the A/P factors, which correlated with the COX-2 expression (R(2) = 0.9496). Although the data support the beneficial antiinflammatory effects from the use of some of the brands tested, others would appear potentially to exacerbate inflammation. To conclude, a ratio based upon the amount and relative proportions of anti- and proinflammatory compounds can be used to predict relative antiinflammatory properties. Also, with access to a diversity of ostensibly similar commercial products, the patient may experience varying therapeutic responses. Finally, current pharmacopoeia monographs, which are generally concerned with a minimum harpagoside content, are inadequate for ensuring the quality of products based on H. procumbens.

Topics: Administration, Cutaneous; Animals; Anti-Inflammatory Agents; Cyclooxygenase 2; Dose-Response Relationship, Drug; Glucosides; Glycosides; Harpagophytum; In Vitro Techniques; Iridoid Glycosides; Phenols; Phytotherapy; Plant Extracts; Pyrans; Skin; Skin Absorption; Swine

Two saponins: scrokoelziside A (1), scrokoelziside B (2), one iridoid glycoside, eurostoside (3), and two flavonoids: nepitrin (4) and homoplantaginin (5), were isolated from the leaves of Scrophularia ningpoensis for the first time. Moreover, eight known compounds: cane sugar (6), harpagide (7), aucubin (8), 6-O-methylcatalpol (9), harpagoside (10), angoroside C (11), beta-sitosterol (12) and beta-sitosterol glucoside (13) were isolated from the roots of S. ningpoensis. Furthermore, the antimicrobial activity of the extracts of the leaves of S. ningpoensis and the 10 compounds (1, 2, 3, 4, 5, 7, 8, 9, 10, 11) was studied in vitro against eight reference strains of bacteria by using the disc-diffusion method and micro-well dilution assay. The extracts of leaves and scrokoelziside A are effective against beta-haemolytic streptococci but had no effect against other strains. The extract of roots and other compounds showed no activity against all bacterial strains at the test concentration.

Topics: Anti-Infective Agents; Bacteria; Flavonoids; Glucosides; Glycosides; Iridoid Glucosides; Iridoid Glycosides; Iridoids; Luteolin; Microbial Sensitivity Tests; Molecular Structure; Plant Extracts; Plant Leaves; Plant Roots; Pyrans; Scrophularia; Sitosterols; Triterpenes

Harpagophytum procumbens, commonly known as Devil's Claw, is indigenous to southern Africa, and extracts of the tubers have been used for centuries in the treatment of a variety of inflammatory disorders. Its major active components, harpagoside (1), harpagide (2), 8-coumaroylharpagide (3), and verbascoside (4), are believed to interact either synergistically or antagonistically in modulating the enzymes responsible for inducing inflammation, although this has not been probed hitherto. In the current work, the ability of these compounds to inhibit the expression of COX-2 following administration to freshly excised porcine skin has been investigated. An ethanol-soluble extract of H. procumbens tubers and two of the pure compounds tested showed promising activity in Western blotting and immunocytochemical assays, with harpagoside (1) and 8-coumaroylharpagide (3) exhibiting greater reductions in COX-2 expression than verbascoside (4). Harpagide (2) caused a significant increase in the levels of COX-2 expression after 6 h of topical application. The data suggest that the efficacy of H. procumbens is dependent upon the ratios of compounds 1-4 present, which is inconsistent with some current official monograph specifications based solely on harpagoside (1) content.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Coumaric Acids; Cyclooxygenase 2 Inhibitors; Epidermis; Glucosides; Glycosides; Harpagophytum; Inflammation; Iridoid Glycosides; Molecular Structure; Phenols; Plants, Medicinal; Pyrans; Swine

Solid-phase extraction cartridges among those usually used for screening in horse doping analyses are tested to optimize the extraction of harpagoside (HS), harpagide (HG), and 8-para-coumaroyl harpagide (8PCHG) from plasma and urine. Extracts are analyzed by liquid chromatography coupled with multi-step tandem mass spectrometry. The extraction process retained for plasma applies BondElut PPL cartridges and provides extraction recoveries between 91% and 93%, with RSD values between 8 and 13% at 0.5 ng/mL. Two different procedures are needed to extract analytes from urine. HS and 8PCHG are extracted using AbsElut Nexus cartridges, with recoveries of 85% and 77%, respectively (RSD between 7% and 19%). The extraction of HG involves the use of two cartridges: BondElut PPL and BondElut C18 HF, with recovery of 75% and RSD between 14% and 19%. The applicability of the extraction methods is determined on authentic equine plasma and urine samples after harpagophytum or harpagoside administration.

Topics: Animals; Chromatography, Liquid; Glycosides; Horses; Hydrogen-Ion Concentration; Iridoid Glycosides; Pyrans; Solid Phase Extraction; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry

A method has been developed for the analysis and characterization in equine urine and plasma of iridoid glycosides: harpagide, harpagoside and 8-para-coumaroyl harpagide, which are the main active principles of Harpagophytum, a plant with antiinflammatory properties. The method involves liquid chromatography coupled with positive electrospray ionization mass spectrometry. The addition of sodium or lithium chloride instead of formic acid in the eluting solvent has been studied in order to enhance the signal and to modify the ion's internal energy. Fragmentation pathways and associated patterns are proposed for each analyte. A comparison of three types of mass spectrometer: a 3D ion trap, a triple quadrupole and a linear ion trap, has been conducted. The 3D ion trap was selected for drug screening analysis whereas the linear ion trap was retained for identification and quantitation analysis.

Topics: Animals; Chromatography, High Pressure Liquid; Glycosides; Harpagophytum; Horses; Iridoid Glycosides; Plant Extracts; Pyrans; Spectrometry, Mass, Electrospray Ionization

To establish a TLC identification method for Xuanshen(Radix Scrophulariae).. Using TLC with harpagoside and harpagide as reference substances.. A TLC identification method of Xuanshen has been established. The specificity of the method has been proved by a comparative detection of Xuanshen from different habitats and several crude drugs which are easily confused with Xuanshen.. The method is simple, accurate and reliable.

Topics: Chromatography, Thin Layer; Drug Contamination; Drugs, Chinese Herbal; Glycosides; Iridoid Glycosides; Pyrans; Scrophularia

The study of the metabolism of iridoid glycosides from Harpagophytum procumbens and Harpagophytum zeyheri by human intestinal bacteria, was realized in order to elucidate compounds responsible for the pharmacological activities of Harpagophytum. Harpagide, harpagoside and 8-O-p-coumaroyl-harpagide were transformed into the pyridine monoterpene alkaloid aucubinine B by human fecal flora and by bacteria isolated from this flora. Aucubinine B was also prepared from harpagide, harpagoside and 8-O-p-coumaroylharpagide, by beta-glucosidase in the presence of NH4+.

Topics: Alkaloids; Bacteria; Coumaric Acids; Glycosides; Humans; Intestines; Iridoid Glycosides; Magnetic Resonance Spectroscopy; Nitrogen; Plants; Pyrans; Pyridines

Effects of the crude methanolic extract of Harpagophytum procumbens secondary roots and two of its active principles, harpagoside and harpagide, on some smooth muscle in vitro have been studied. The results obtained show how the action of H. procumbens is due to a complex interaction between the various active principles contained in the drug and suggest that they, especially harpagoside, interfere with the mechanisms that regulate the influx of calcium in the cells.

Topics: Acetylcholine; Animals; Barium; Barium Compounds; Chlorides; Glycosides; Guinea Pigs; Ileum; In Vitro Techniques; Iridoid Glycosides; Jejunum; Male; Muscle Contraction; Muscle, Smooth; Plant Extracts; Pyrans; Rabbits

In conscious normotensive rats the dried crude methanolic extract of Harpagophytum procumbens secondary roots caused a significant dose-dependent reduction of arterial blood pressure. The decrease was significant only at higher doses given by gavage (dried extract = 400 mg/kg). At the same time a decrease of heart rate was observed. In the same experimental conditions, harpagoside presented an activity lower than doses of Harpagophytum procumbens extract containing corresponding quantities of harpagoside. In spontaneously beating Langendorff preparations of rabbit heart, the Harpagophytum procumbens methanolic extract caused a mild decrease in the heart rate with a concomitant mild positive inotropic effect at lower doses but a marked negative inotropic effect at higher doses. The coronary flow decreased at higher doses only. The negative chronotropic and positive inotropic effects of harpagoside were comparatively higher with respect to that of the extract, whereas harpagide had only a slight negative chronotropic effect and a considerable negative inotropic one. Both in experiments on intact rats and on isolated rabbit heart, the Harpagophytum procumbens extract also demonstrated a protective action with regard to arrhythmias induced by aconitine, and particularly to those provoked by calcium chloride and epinephrine--chloroform.

Topics: Animals; Anti-Arrhythmia Agents; Blood Pressure; Electrocardiography; Glycosides; Hemodynamics; In Vitro Techniques; Iridoid Glycosides; Lidocaine; Male; Myocardial Contraction; Plant Extracts; Plants, Medicinal; Pyrans; Rabbits; Rats; Rats, Inbred Strains