Research update of St. John's Wort Hypericum perforatum and hypericin.

  seminal trace...St. John's Wort Extract.Hypericins 0.3%1%UV.Hyperforin 0.3%HPLC.Hypericin.M.F.C30H16O8.M.W.504.45.CAS.No.548-04-9.EINECS.No.208-941-0.Acute toxicity of hyperin.Hyperforin.0.3%HPLC.M.F.C35H52O4.M.W.536.80.CAS No.11079-53-1.Hypericum perforatum Extract...

 


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   Research update of St. John's Wort Hypericum perforatum and hypericin.

   A Mechanistic Study on Altered Pharmacokinetics of Irinotecan by St. John's Wort.:Curr Drug Metab. 2007 Feb;8(2):157-71.Hu ZP, Yang XX, Chen X, Cao J, Chan E, Duan W, Huang M, Yu XQ, Wen JY, Zhou SF.Department of Pharmacy, Faculty of Science, National University of Singapore, Science Drive 4, 117543, Singapore; shufengzhou2006@hotmail.com.[PMID: 17305494]

 Irinotecan (CPT-11) is an important anticancer drug in management of advanced colon cancer. A marked protective effect on CPT-11-induced blood and gastrointestinal toxicity is obtained by combination of St. John's wort (SJW) in recent clinical and rat studies. However, the mechanism is unclear. This study aimed to explore the effects of SJW on the pharmacokinetics of CPT-11 and its major metabolites (SN-38 and SN-38 glucuronide) in rats and the underlying mechanisms using several in vitro models. Short-term (3 days) and long-term (14 days) pretreatment with SJW were conducted in rats to examine the effects of co-administered SJW on the plasma pharmacokinetics of CPT-11, SN-38 and SN-38 glucuronide. Rat liver microsomes and a rat hepatoma cell line, H4-II-E cells, were utilized to study the effects of aqueous and ethanolic extracts (AE and EE) and major active components (hyperforin, hypericin and quercetin) of SJW on CPT-11 and SN-38 metabolism and intracellular accumulation. Co-administered SJW for consecutive 14 days significantly decreased the initial plasma concentration (C(0)) of CPT-11, the area under the concentration-time curve (AUC(0-10hr)) and maximum plasma concentration (C(max)) of SN-38. The ethanolic extracts (EE) of SJW at 5 microg/ml significantly decreased SN-38 glucuronidation by 45% (P < 0.05) in rat hepatic microsomes. Pre-incubation of aqueous SJW extracts (AE) at 10 microg/ml, SJW EE at 5 microg/ml, and quercetin at 10 microM significantly increased the glucuronidation of SN-38 in H4-II-E cells. A 2-hr pre-incubation of quercetin (100 microM) significantly increased the intracellular accumulation of CPT-11 (P < 0.05). However, pre-incubation of hypericin (20 nM and 200 nM) and hyperforin (1microM) significantly decreased the intracellular accumulation of CPT-11. In addition, pre-incubation of hypericin, SJW EE and quercetin significantly increased the intracellular accumulation of SN-38. Aqueous and ethanolic SJW extracts and its major active components did not alter the plasma protein binding of CPT-11 and SN-38. These results indicated that the aqueous and ethanolic extracts of SJW and its major active components could markedly alter glucuronidation of SN-38 and intracellular accumulation of CPT-11 and SN-38, which probably provides partial explanation for the altered plasma pharmacokinetics of CPT-11 and SN-38 and the antagonizing effects on the toxicities of CPT-11. Further studies are needed to explore the role of both pharmacokinetic and pharmacodynamic components in the protective effect of SJW against the toxicities of CPT-11.

   The production of hypericins in two selected Hypericum perforatum shoot cultures is related to differences in black gland structure.:Plant Physiol Biochem. 2006 Dec 28;Kornfeld A, Kaufman PB, Lu CR, Gibson DM, Bolling SF, Warber SL, Chang SC, Kirakosyan A.Department of Biological Sciences, Humboldt State University, Arcata, CA 95521, USA.[PMID: 17300946]

 In vitro shoot cultures of Hypericum perforatum derived from wild populations grown in Armenia have a wide variation of hypericin and pseudohypericin metabolite content. We found that a germ line denoted as HP3 produces six times more hypericin and fourteen times more pseudohypericin than a second line labeled HP1. We undertook a structural comparison of the two lines (HP1 and HP3) in order to see if there are any anatomical or morphological differences that could explain the differences in production of these economically important metabolites. Analysis by LM (light microscopy), SEM (scanning electron microscopy), and TEM (transmission electron microscopy) reveals that the hypericin/pseudohypericin-containing black glands located along the margins of the leaves consist of a peripheral sheath of flattened cells surrounding a core of interior cells that are typically dead at maturity. The peripheral cells of the HP3 glands appear less flattened than those of the HP1 glands. This may indicate that the peripheral cells are involved in hypericin/pseudohypericin production. Furthermore, we find that these peripheral cells undergo a developmental transition into the gland's interior cells. The fact that the size of the peripheral cells may correlate with metabolite production adds a new hypothesis for the actual site of hypericin synthesis.

   Quantitative phytochemical analyses of six hypericum species growing in slovenia.:Planta Med. 1999 May;65(4):388-90.Umek A, Kreft S, Kartnig T, Heydel B.Faculty of Pharmacy, University of Ljubljana, Ljubljana, Slovenia.[PMID: 17260265]

 The 74 samples of six HYPERICUM species (H. PERFOROTUM, H. HIRSUTUM, H. MACULATUM, H. TETRAPTERUM, H. MONTANUM, AND H. HUMIFUSUM) were collected around Slovenia and analysed for the content of ten substances (rutin, hyperoside, isoquercetin, quercitrin, quercetin, I3,II8-biapigenin, amentoflavone, pseudohypericin, hypericin, and hyperforin). The flowers were analysed separately from the green parts of the plants (herbs). The highest content of most of the substances was found in the flowers of H. PERFOROTUM. Among the herbal samples (without flowers), H. MONTANUM and H. HIRSUTUM contained significantly higher levels of amentoflavone (average 3-fold and 2.5-fold higher, respectively), than the herbs of H. PERFOROTUM. In the herbal part of H. PERFOROTUM the contents of all constituents strongly correlate with the contents of the same compound in flowers, except for the content of amentoflavone, which is independent in these two parts. Rutin and hyperoside are in positive correlation, and quercitrin is in negative correlation with the altitude of the growing site.

   Identification of the major constituents of Hypericum perforatum by LC/SPE/NMR and/or LC/MS.:Phytochemistry. 2007 Feb;68(3):383-93. Epub 2006 Dec 29.Tatsis EC, Boeren S, Exarchou V, Troganis AN, Vervoort J, Gerothanassis IP.Section of Organic Chemistry and Biochemistry, Department of Chemistry, University of Ioannina, University Campus, Ioannina, Epirus GR-45110, Greece.[PMID: 17196625]

 The newly established hyphenated instrumentation of LC/DAD/SPE/NMR and LC/UV/(ESI)MS techniques have been applied for separation and structure verification of the major known constituents present in Greek Hypericum perforatum extracts. The chromatographic separation was performed on a C18 column. Acetonitrile-water was used as a mobile phase. For the on-line NMR detection, the analytes eluted from column were trapped one by one onto separate SPE cartridges, and hereafter transported into the NMR flow-cell. LC/DAD/SPE/NMR and LC/UV/MS allowed the characterization of constituents of Greek H. perforatum, mainly naphtodianthrones (hypericin, pseudohypericin, protohypericin, protopseudohypericin), phloroglucinols (hyperforin, adhyperforin), flavonoids (quercetin, quercitrin, isoquercitrin, hyperoside, astilbin, miquelianin, I3,II8-biapigenin) and phenolic acids (chlorogenic acid, 3-O-coumaroylquinic acid). Two phloroglucinols (hyperfirin and adhyperfirin) were detected for the first time, which have been previously reported to be precursors in the biosynthesis of hyperforin and adhyperforin.

   Antiinflammatory activity of ethanol extracts of Hypericum perforatum L., H. barbatum Jacq., H. hirsutum L., H. richeri Vill. and H. androsaemum L. in rats.:Phytother Res. 2007 Feb;21(2):176-80.Savikin K, Dobric S, Tadic V, Zdunic G. Institute for Medicinal Plants Research, Tadeusa Koscuska 1, 11000 Belgrade, Serbia and Montenegro. ksavikin@iplb.co.yu.[PMID: 17143926]

 Ethanol extracts of the aerial parts of six Hypericum species, H. barbatum, H. androsaemum, H. richerii, H. hirsutum, H. perforatum and H. perforatum cultivated on mountain Tara were tested for antiinflammatory activity in comparison with indomethacin (IND) using the carrageenan-induced rat paw edema test. It was found that all examined extracts produced antiinflammatory activity, particularly those from H. hirsutum, and both wild and cultivated H. perforatum. A dose-dependent antiinflammatory effect was especially pronounced in extracts showing some lower antiinflammatory activity. There was no correlation between the amount of hypericin in the extracts and their antiinflammatory activity.


   Plant-environment interactions: Accumulation of hypericin in dark glands of Hypericum perforatum.:Ann Bot (Lond). 2006 Oct;98(4):793-804. Epub 2006 Aug 4.

 BACKGROUND AND AIMS: Hypericum perforatum is a perennial herbaceous plant and an extract from this plant has a significant antidepressant effect when administered to humans. The plant is characterized by its secretory glands, also known as dark glands, which are mainly visible on leaves and flowers. The current study evaluates the influence of several environmental factors and developmental stages of the plant on the accumulation and synthesis of hypericin and pseudohypericin (Hy-G), the major bioactive constituents, in H. perforatum plants. METHODS: The appearance of dark glands on different parts of the plant, under several environmental conditions, was monitored by microscopy. Hy-G concentrations were quantified by high-performance liquid chromatography. KEY RESULTS: A significant presence of dark glands accompanying the highest concentrations of Hy-G was observed in the stamen tissues more than in any other organ of H. perforatum. A linear relationship between the number of dark glands and net photosynthetic rate of the leaf and Hy-G concentration in the leaf tissue was also established. A very high concentration of Hy-G was measured in the dark-gland tissues, but in the tissues without any dark glands it was almost absent. The presence of emodin, a precursor of Hy-G, at a high concentration in the dark-gland tissues, and its absence in the surrounding tissues was also observed, suggesting that the site of biosynthesis of Hy-G is in the dark-gland cells. A significantly low concentration of Hy-G (occasionally non-detectable) was measured in the xylem sap of the stem tissues. The dark-gland tissues collected from leaves, stems or flowers contained similar concentrations of Hy-G. CONCLUSIONS: The concentration of Hy-G in various organs of H. perforatum plants is dependent on the number of dark glands, their size or area, not on the location of the dark glands on the plant. The study provides the first experimental evidence that Hy-G is synthesized and accumulates in dark glands.

   Reduction in hypericin-induced phototoxicity by Hypericum perforatum extracts and pure compounds.:J Photochem Photobiol B. 2006 Nov 1;85(2):118-30. Epub 2006 Jul 21.Schmitt LA, Liu Y, Murphy PA, Petrich JW, Dixon PM, Birt DF.The Center for Research on Dietary Botanical Supplements, Iowa State University, 215 MacKay Building, Ames, IA 50011-1120, USA.[PMID: 16859921]

 Clinical evidence suggests that administration of Hypericum perforatum (Hp) extracts containing the photo-activated hypericin compounds may cause fewer skin photosensitization reactions than administration of pure hypericin. This study was conducted to determine whether the phototoxicity of hypericin in HaCaT keratinocytes could be attenuated by H. perforatum extracts and constituents. Two extracts, when supplemented with 20 microM hypericin: (1) an ethanol re-extraction of residue following a chloroform extraction (denoted ethanol(-chloroform)) (3.35 microM hypericin and 124.0 microM total flavonoids); and (2) a chloroform extract (hypericin and flavonoids not detected), showed 25% and 50% (p<0.0001) less phototoxicity than 20 microM hypericin alone. Two H. perforatum constituents, when supplemented with 20 microM hypericin: (1) 10 microM chlorogenic acid; and (2) 0.25 microM pyropheophorbide, exhibited 24% (p<0.05) and 40% (p<0.05) less phototoxicity than 20 microM hypericin alone. The peroxidation of arachidonic acid was assessed as a measure of oxidative damage by photo-activated hypericin, but this parameter of lipid peroxidation was not influenced by the extracts or constituents. However alpha-tocopherol, a known antioxidant also did not influence the amount of lipid peroxidation induced in this system. These observations indicate that hypericin combined with H. perforatum extracts or constituents may exert less phototoxicity than pure hypericin, but possibly not through a reduction in arachidonic acid peroxidation.

   Hypericum perforatum: a 'modern' herbal antidepressant: pharmacokinetics of active ingredients.:Clin Pharmacokinet. 2006;45(5):449-68.Wurglics M, Schubert-Zsilavecz M.Institute of Pharmaceutical Chemistry, ZAFES, J.W. Goethe University, Frankfurt am Main, Germany. Wurglics@pharmchem.uni-frankfurt.de.[PMID: 16640452]

 Hypericum perforatum (St John's Wort [SJW]) counts among the most favourite herbal drugs, and is the only herbal alternative to classic synthetic antidepressants in the therapy of mild to moderate depression. Several clinical studies have been conducted to verify the effectiveness of ethanolic or methanolic extracts of SJW. Alcoholic SJW extracts are a mixture of substances with widely varying physical and chemical properties and activities. Hyperforin, a phloroglucinol derivative, is the main source of pharmacological effects caused by the consumption of alcoholic extracts of SJW in the therapy of depression. However, several studies indicate that flavone derivatives, e.g. rutin, and also the naphthodianthrones hypericin and pseudohypericin, take part in the antidepressant efficacy. In contrast to the amount of documentation concerning clinical efficacy, oral bioavailability and pharmacokinetic data about the active components are rather scarce. The hyperforin plasma concentration in humans was investigated in a small number of studies. The results of these studies indicate a relevant plasma concentration, comparable with that used in in vitro tests. Furthermore, hyperforin is the only ingredient of H. perforatum that could be determined in the brain of rodents after oral administration of alcoholic extracts. The plasma concentrations of the hypericins were, compared with hyperforin, only one-tenth and, until now, the hypericins could not be found in the brain after oral administration of alcoholic H. perforatum extracts or pure hypericin. Until now, the pharmacokinetic profile of the flavonoids in humans after oral administration of an alcoholic H. perforatum extract has been investigated in only one study. More data are available for rutin and the aglycone quercetin after administration of pure substances or other flavonoid sources.

   Investigation of the effect on photosensitivity following multiple oral dosing of two different hypericum extracts in healthy men.:Arzneimittelforschung. 2006;56(3):212-21.Schulz HU, Schurer M, Bassler D, Weiser D.LAFAA Laboratory for Contract Research in Clinical Pharmacology and Biopharmaceutical Analytics GmbH, Bad Schwartau, Germany. lafaa_gmbh@t-online.de.[PMID: 16618014]

 The naphthodiantrones hypericin and pseudohypericin, ingredients of hypericum extracts, are known as potent photosensitizers that may cause phototoxic effects in grazing animals after excessive ingestion of hypericum species and in some cases in higher concentrations of hypericum extracts oder pure hypericin in humans as well. Therefore, the objective of the present studies was to investigate the effect of two different hypericum extracts (STW 3, STW 3-VI) on photosensitivity with respect to minimal erythema dose (MED) after 14 days treatment. Both open, multiple-dose, one-phase studies were conducted in 20 healthy men, receiving one tablet per day. MED values were determined prior to hypericum extract administration (baseline) and after 14 days treatment using an erythem tester emitting a light very similar to sun light (main emission spectrum: 285-350 nm). Skin reactions with respect to MED were evaluated 12 h, 24 h (primary endpoint), 48 h and 7 days after irradiation. All volunteers reached steady-state of hypericin/pseudohypericin plasma concentrations before study day 14, when the irradiation under treatment conditions took place. In all subjects MED was measurable under baseline and under hypericum treatment conditions. With respect to the primary endpoint, in both studies, mean MED (24 h) were not significantly different between baseline and after 14 days hypericum treatment. However, individually photosensitivity of the skin could increase under treatment conditions, just as well photosensitivity could decrease or remain unchanged. There were no clinically relevant changes in the laboratory parameters, the vital signs, physical findings and other observations related to safety during the examinations. In one study (STW 3), two adverse events were reported, both described as hypersensitivity to light in mild Intensity. The two studies showed that treatment with the two hypericum extracts under steady state and under prescribed conditions were safe medications without significant increases of photosensitivity.

   Evaluation of the light-sensitive cytotoxicity of Hypericum perforatum extracts, fractions, and pure compounds.:J Agric Food Chem. 2006 Apr 19;54(8):2881-90.Schmitt LA, Liu Y, Murphy PA, Birt DF.The Center for Research on Dietary Botanical Supplements, Iowa State University, Ames, Iowa 50014, USA.[PMID: 16608204]

 Hypericum perforatum (Hp) is known for possessing antidepressant and antiviral activities. Despite its use as an alternative to conventional antidepressants, the identification of the cytotoxic chemicals derived from this herb is incomplete. In this study, the cytotoxicity of Hp extracts prepared in solvents ranging in polarity, fractions of one extract, and purified compounds were examined in three cell lines. All extracts exhibited significant cytotoxicity; those prepared in ethanol (no hyperforin, 3.6 microM hypericin, and 134.6 microM flavonoids) showed between 7.7 and 77.4% cell survival (p < 0.0001 and 0.01), whereas the chloroform and hexane extracts (hyperforin, hypericin, and flavonoids not detected) showed approximately 9.0 (p < 0.0001) and 4.0% (p < 0.0001) survival. Light-sensitive toxicity was observed primarily with the ethanol extracts sequentially extracted following removal of material extracted in either chloroform or hexane. The absence of light-sensitive toxicity with the Hp extracts suggests that the hypericins were not playing a prominent role in the toxicity of the extracts.


   Comparison of methods for the exhaustive extraction of hypericins, flavonoids, and hyperforin from Hypericum perforatum L.:J Agric Food Chem. 2006 Apr 5;54(7):2750-3.Smelcerovic A, Spiteller M, Zuehlke S.Institute of Environmental Research (INFU), University of Dortmund, Otto-Hahn-Strasse 6, D-44221 Dortmund, Germany.[PMID: 16569071]

 Renewed interest in plant-derived drugs has led to an increased need for efficient extraction methods. Hypericum perforatum L. contains several groups of bioactive compounds with noteworthy pharmacological activities. Direct sonication of H. perforatum was investigated and compared with conventional maceration, indirect sonication, Soxhlet extraction, and accelerated solvent extraction (ASE). Highly selective liquid chromatography/tandem mass spectrometry analysis showed that the content of six investigated active compounds (hypericin, pseudohypericin, hyperoside, rutin, quercitrin, and hyperforin) in extracts obtained by direct sonication was significantly higher than in extracts obtained by the other methods. The active compound contents increased on increasing the ultrasonic power from 40 to 60 W when using direct sonication. Conventional maceration gave the lowest amount of analyzed active compounds. Soxhlet extraction gave better results than ASE or indirect sonication.

   Induction of hypericins in Hypericum perforatum in response to chromium.:Fitoterapia. 2006 Apr;77(3):164-70. Epub 2006 Mar 6.Tirillini B, Ricci A, Pintore G, Chessa M, Sighinolfi S. Institute of Botany, University of Urbino, Via Bramante, 28, 61029 Urbino, Italy. tirillini@uniurb.it.[PMID: 16554124]

 Seedlings of Hypericum perforatum were grown with 0.01 and 0.1 mM of chromium added to the nutrient media. A treatment with 0.01 mM Cr(VI) for seven days resulted in an increased production of protopseudohypericin (+135%), hypericin (+38%) and pseudohypericin (+5%). Treatment with 0.1 mM Cr(VI) for two days also caused an increase of protopseudohypericin (+167%), hypericin (25%) and pseudohypericin (+5%). The greatest effect of chromium treatment was observed at a concentration of 0.1 mM for seven days: protopseudohypericin increased +404% and pseudohypericin to +379%. Hypericin was not affected by this treatment.

   Development and evaluation of methods for determination of naphthodianthrones and flavonoids in St. John's wort.:J Chromatogr A. 2006 May 19;1115(1-2):93-102. Epub 2006 Mar 22.Williams FB, Sander LC, Wise SA, Girard J. National Institute of Standards and Technology, Gaithersburg, MD 20899, USA.[PMID: 16554056]

 Several major constituents in St. John's wort were determined for a homogenized plant sample. Three extraction techniques were evaluated: Soxhlet extraction, pressurized-fluid extraction (PFE), and sonication extraction. Levels of nine constituents (chlorogenic acid, rutin, hyperoside, isoquercitrin, quercitrin, quercetin, amentoflavone, pseudohypericin, and hypericin) were measured using liquid chromatography with ultraviolet/visible absorbance, mass spectrometric, and fluorescence detection. Levels of total naphthodianthrones determined by liquid chromatography (LC) with absorbance detection at 590 nm were compared with levels determined by direct spectrophotometry at the same wavelength. Additionally, the methods described in this paper were applied to several brands of St. John's wort finished products.

   Photophysical properties of Hypericum perforatum L. extracts--novel photosensitizers for PDT.:J Photochem Photobiol B. 2006 Feb 1;82(2):146-51. Epub 2006 Jan 4.Skalkos D, Gioti E, Stalikas CD, Meyer H, Papazoglou TG, Filippidis G. Department of Materials Science and Engineering, University of Ioannina, Ioannina GR-45110, Greece. dskalkos@cc.uoi.gr.[PMID: 16388961]

 We report the preparation of the methanolic extract (ME), and polar methanolic fraction (PMF) from the plant Hypericum perforatum L. The extracts contain various photosensitizing constituents such as naphthodianthrone derivatives (in 1.37% w/w), and chlorophylls (in 0.08% w/w). Upon light emission these constituents can be activated, providing photodynamic properties to the extracts, and making them a potent, new class, natural photosensitizers for use in photodynamic therapy (PDT), and photodynamic diagnosis (PDD). The absorbance spectra of the extracts are similar to the spectrum of hypericin, the main naphthodianthrone identified within, with two major bands at 548 and 590 nm. The fluorescence spectra in ethanol exhibit two main bands around 595 and 640 nm, in accordance with the spectrum of pure hypericin. The fluorescence intensity of PMF at 595 nm is only eight times less than the intensity of pure hypericin at the same wavelength, even though its hypericin concentration is only 0.57% w/w. The dependence of the PMF fluorescence signal on the pH of the medium, alone and in comparison with the signal of hypericin, has been investigated. PMF signal fades steadily, and smoothly both in acidic, and basic environment.

   Antidepressant-like components of Hypericum perforatum extracts: an overview of their pharmacokinetics and metabolism.:Curr Drug Metab. 2005 Dec;6(6):531-43.Caccia S.Istituto di Ricerche Farmacologiche "Mario Negri", via Eritrea 62, 20157 Milan, Italy. caccia@ marionegri.it.[PMID: 16379667]

 Extracts of Hypericum perforatum are becoming increasingly popular for the treatment of mild to moderate depression, despite the lack of consensus on their efficacy. Although the mechanism(s) of this action are still debated, several components, including the naphthodianthrones hypericin and pseudohypericin, the acylphloroglucinol hyperforin and some flavonols, are believed to play major roles in the antidepressant-like effects. Some of these also increase the expression of the P-glycoprotein transporter and others the expression of cytochrome P450 enzymes, possibly contributing to the interactions involving the extracts and conventional drugs. However, few pharmacokinetic studies of naphthodianthrones and hyperforin have appeared and none has yet evaluated the exposure to unchanged quercetin and its glycosides after intake of extracts. There are no formal pharmacokinetic studies in special populations. Bioavailability appears low, giving variable steady-state plasma concentrations, whose prediction may be complicated by non-linearity for hypericin and hyperforin. Data on tissue distribution are scarce, and it appears that hypericin and hyperforin do not reach the central nervous system in appreciable concentrations in animals. Clearance is low-intermediate, with little or no unchanged compounds excreted with urine. Although some potentially active conjugated metabolites have been identified for quercetin and its glycosides after intake of authentic compounds or flavonol-rich foods, these too have been characterised little with regard to their pharmacokinetics and central activities. Thus, further pharmacokinetic and pharmacodynamic studies of the main components and their metabolites are urgently needed to clarify the role of each constituent and provide more rational and safe regimens for people preferring "natural" drugs.


   Approach to novel functional foods for stress control 4. Regulation of serotonin transporter by food factors.:[PMID: 16366509]

 Serotonin transporters (SERTs) are pre-synaptic proteins specialized for the clearance of serotonin following vesicular release at central nervous system (CNS) and enteric nervous system synapses. SERTs are high affinity targets in vivo for antidepressants such as serotonin selective reuptake inhibitors (SSRIs). These include 'medical' psychopharmacological agents such as analgesics and antihistamines, a plant extract called St John's Wort (Hypericum). Osteoclasts are the primary cells responsible for bone resorption. They arise by the differentiation of osteoclast precursors of the monocyte/macrophage lineage. The expression of SERTs was increased in RANKL-induced osteoclast-like cells. Using RANKL stimulation of RAW264.7 cells as a model system for osteoclast differentiation, we studied the direct effects of food factor on serotonin uptake. The SSRIs (fluoxetine and fluvoxamine) inhibited markedly (approximately 95%) in serotonin transport in differentiated osteoclast cells. The major components of St. John's Wort, hyperforin and hypericine were significantly decreased in serotonin transport activity. Thus, a new in vitro model using RANKL-induced osteoclast-like cells may be useful to analyze the regulation of SERT by food factors and SSRIs.

   Quantitative analysis of the major constituents of St John's wort with HPLC-ESI-MS.:J Pharm Pharmacol. 2005 Dec;57(12):1645-52.Chandrasekera DH, Welham KJ, Ashton D, Middleton R, Heinrich M. Centre for Pharmacognosy and Phytotherapy, School of Pharmacy, University of London, 29-39 Brunswick Square, London WC1N 1AX, UK.[PMID:16354409]

 A method was developed to profile the major constituents of St John's wort extracts using high-performance liquid chromatography-electrospray mass spectrometry (HPLC-ESI-MS). The objective was to simultaneously separate, identify and quantify hyperforin, hypericin, pseudohypericin, rutin, hyperoside, isoquercetrin, quercitrin and chlorogenic acid using HPLC-MS. Quantification was performed using an external standardisation method with reference standards. The method consisted of two protocols: one for the analysis of flavonoids and glycosides and the other for the analysis of the more lipophilic hypericins and hyperforin. Both protocols used a reverse phase Luna phenyl hexyl column. The separation of the flavonoids and glycosides was achieved within 35 min and that of the hypericins and hyperforin within 9 min. The linear response range in ESI-MS was established for each compound and all had linear regression coefficient values greater than 0.97. Both protocols proved to be very specific for the constituents analysed. MS analysis showed no other signals within the analyte peaks. The method was robust and applicable to alcoholic tinctures, tablet/capsule extracts in various solvents and herb extracts. The method was applied to evaluate the phytopharmaceutical quality of St John's wort preparations available in the UK in order to test the method and investigate if they contain at least the main constituents and at what concentrations.

   The extent of induction of CYP3A by St. John's wort varies among products and is linked to hyperforin dose.:Eur J Clin Pharmacol. 2006 Jan;62(1):29-36. Epub 2005 Dec 10.Mueller SC, Majcher-Peszynska J, Uehleke B, Klammt S, Mundkowski RG, Miekisch W, Sievers H, Bauer S, Frank B, Kundt G, Drewelow B. Center of Pharmacology and Toxicology, Institute of Clinical Pharmacology, University of Rostock, Schillingallee 70, 18057, Rostock, Germany. silke.mueller@med.uni-rostock.de.[PMID: 16341856]

 OBJECTIVE: Induction of CYP3A by St. John's wort (SJW) extracts with high hyperforin (HYF) content is well described. Since SJW products vary in the amount of HYF and other main constituents, the aim of the study was to evaluate the effect on CYP3A function of SJW preparations with a range from very low to high HYF content. METHODS: Forty-two male, healthy volunteers were randomized into six parallel SJW medication groups with varying composition especially with regard to HYF content. Midazolam plasma concentration profiles were characterized after a single oral dose of 7.5 mg midazolam on the day before and on the 14th day of SJW medication. RESULTS: All SJW preparations tested resulted in a decrease in midazolam AUC, although the extent of the effect differed. The extract LI 160 (HYF 41 mg/day) decreased midazolam AUC0-12h by 79.4% (95% CI -88.6; -70.1), which was significantly greater than the effect by any other medication (p<0.05). SJW powder tablets 2.7 g/day (HYF 12 mg/day) resulted in a midazolam AUC0-12h decrease of 47.9% (95% CI -59.7;-36.2), while 2.7 g/day SJW powder tablets that were almost devoid of HYF (0.13 mg/day) reduced midazolam AUC0-12h by only 21.1% (95% CI -33.9; -8.3). Considering all six SJW medications tested, the extent of midazolam AUC decrease correlated significantly with increasing HYF dose (r=-0.765, p<0.001), but not with hypericin dose (r=-0.067; p=0.673). CONCLUSION: The extent of induction of CYP3A varies among St. John's wort products and depends on hyperforin dose.

   The lipophilic extract of Hypericum perforatum exerts significant cytotoxic activity against T24 and NBT-II urinary bladder tumor cells.:Planta Med. 2005 Nov;71(11):1030-5.Skalkos D, Stavropoulos NE, Tsimaris I, Gioti E, Stalikas CD, Nseyo UO, Ioachim E, Agnantis NJ.Department of Material Sciences & Engineering, University of Ioannina, Greece. dskalkos@cc.uoi.gr.[PMID: 16320204]

 Hypericum perforatum L. (St. John's wort) is a medicinal plant used for many pathologies, especially for the treatment of mild to moderate depression. In the present study we have investigated the cytotoxic activity of the locally collected (Epirus region) Hypericum perforatum L. against cultured T24 and NBT-II bladder cancer cell lines. The lipophilic extract of the herb, prepared using petroleum ether, induced apoptosis displaying LC(50) values at concentrations as low as 4 and 5 microg/mL. A fraction of this extract displayed 60 % cell growth inhibition at a concentration of 0.95 microg/mL. Evaluating the importance of various biologically active components of the extract, it was found that hypericins (hypericin, pseudohypericin, etc.) were identified only in the methanolic (lipophobic) extract of the herb, and not in the active lipophilic extract. In addition, hyperforin concentrations in the lipophilic extract and its most active fraction, were 0.94 microg/mL, and 0.17 microg/mL, respectively, while the active cytotoxic concentration of pure hyperforin appeared in the range of 1.8 microg/mL - 5.0 microg/mL. Therefore, pure hyperforin does not seem to contribute significantly to the cytotoxicity activity. Chlorophylls were identified in low, not significantly different, concentrations in all extracts and fractions and were not correlated to the biological activity. Owing to the combination of significant cytotoxic activity, natural abundance and low toxicity, the lipophilic extract of Hypericum perforatum holds the promise of being an interesting, new, antiproliferative agent against bladder cancer that deserves further investigation.

   Content analysis and stability evaluation of selected commercial preparations of St. John's wort.:Drug Dev Ind Pharm. 2005 Oct;31(9):907-16.Shah AK, Avery BA, Wyandt CM. Department of Pharmaceutics, School of Pharmacy, University of Mississippi, University, Mississippi 38677, USA. ashah@olemiss.edu.[PMID: 16306003]

 Content analysis and stability studies were performed for the commercial products of St. John's wort. Six marketed formulations were analyzed for their hypericin and pseudohypericin content. These products were standardized to contain 0.3% hypericin. Results revealed total hypericin as 7.72-38.57% of the labeled claim with varying concentrations of pseudohypericin. Stability studies were carried out under three different storage conditions: 1) 25+/-2 degrees C, 60+/-5%RH for six months, 2) 40+/-2 degrees C, 75+/-5%RH for six months, and 3) 50 degrees C for one month. Tablet formulations were also analyzed for their hardness and friability. Stability studies revealed significant decrease in the content of the marker compounds with time.


   Investigation of pharmacokinetic data of hypericin, pseudohypericin, hyperforin and the flavonoids quercetin and isorhamnetin revealed from single and multiple oral dose studies with a hypericum extract containing tablet in healthy male volunteers.:Arzneimittelforschung. 2005;55(10):561-8.Schulz HU, Schurer M, Bassler D, Weiser D.LAFAA Laboratory for Contract Research in Clinical Pharmacology and Biopharmaceutical Analytics GmbH, Bad Schwartau, Germany. lafaa_gmbh@t-online.de.[PMID: 16294501]

 Hypericins, hyperforin and flavonoids are discussed as the main components contributing to the antidepressant action of St. John's wort (Hypericum perforatum). Therefore, the objective of the two open phase I clinical trials was to obtain pharmacokinetic data of these constituents from a hypericum extract containing tablet: hypericin, pseudohypericin, hyperforin, the flavonoid aglycone quercetin, and its methylated form isorhamnetin. Each trial included 18 healthy male volunteers who received the test preparation, containing 900 mg dry extract of St John's wort (STW 3-VI, Laif 900), either as a single oral dose or as a multiple once daily dose over a period of 14 days. Concentration/time curves were determined for the five constituents, for 48 h after single dosing and for 24 h on day 14 at the end of 2 weeks of continuous daily dosing. After single dose intake, the key pharmacokinetic parameters were determined as follows: Hypericin: Area under the curve (AUC(0-infinity)) = 78.33 h x ng/ml, maximum plasma concentration (Cmax) = 3.8 ng/ml, time to reach Cmax (tmax) = 7.9 h, and elimination half-life (t1/2) = 18.71 h; pseudohypericin: AUC(0-infinity) = 97.28 h x ng/ml, Cmax = 10.2 ng/ml, tmax = 2.7 h, t1/2 = 17.19 h; hyperforin: AUC(0-infinity) = 1550.4 h x ng/ml, Cmax = 122.0 ng/ml, tmax = 4.5 h, t1/2 = 17.47 h. Quercetin and isorhamnetin showed two peaks of maximum plasma concentration separated by about 3-3.5 h. Quercetin: AUC(0-infinity) = 417.38 h x ng/ml, Cmax (1) = 89.5 ng/ml, tmax (1) = 1.0 h, Cma (2) = 79.1 ng/ml, tmax (2) = 4.4 h, t1/2 = 2.6 h; isorhamnetin: AUC(0-infinity) = 155.72 h x ng/ml, Cmax (1) = 12.5 ng/ml, tmax (1) = 1.4 h, Cmax (2) = 14.6 ng/ml, tmax (2) = 4.5 h, t1/2 = 5.61 h. Under steady state conditions reached during multiple dose administration similar results were obtained. Further pharmacokinetic characteristics calculated from the obtained data were the mean residence time (MRT), the lag-time, the peak-trough fluctuation (PTF), the lowest observed plasma concentration (Cmin), and the average plasma concentration (Cav). The data obtained for the five consitituents generally corresponded well with values previously published. The trial preparation was well tolerated.

   Photoinduced antitumour effect of hypericin can be enhanced by fractionated dosing.:Phytomedicine. 2005 Sep;12(9):680-3.Cavarga I, Brezani P, Fedorocko P, Miskovsky P, Bobrov N, Longauer F, Rybarova S, Mirossay L, Stubna J.Medical School Hospital of L. Pasteur, Kosice, Slovakia.[PMID: 16194057]

 The in vivo antitumour activity of the natural photosensitizer hypericin was evaluated. C3H/DiSn mice were inoculated with fibrosarcoma G5:1:13 cells. When the tumour reached a volume of 40-80mm3 the mice were intraperitoneally injected with hypericin, either in a single dose (5mg/kg; 1 or 6h before laser irradiation) or two fractionated doses (2.5 mg/kg; 6 and 1 h before irradiation with laser light; 532 nm, 70mW/cm2, 168 J/cm2). All tumours in control groups treated with hypericin alone as well as those irradiated with laser light alone had similar growth rates and none of these tumours regressed spontaneously. Complete remission of tumour in photodynamic therapy (PDT)-treated groups was similar (14-17% single dose vs. 33% fractionated dose), but the fractionated schedule of hypericin dosing was found to be more efficient than the single dose, measured by survival assay (p < 0.05). Our experimental model showed that fractionated administration of hypericin can produce a better therapeutic response than single administration.

   Identification and quantification of hypericin and pseudohypericin in different Hypericum perforatum L. in vitro cultures.:Plant Physiol Biochem. 2005 Jun;43(6):591-601.Gadzovska S, Maury S, Ounnar S, Righezza M, Kascakova S, Refregiers M, Spasenoski M, Joseph C, Hagege D. Laboratoire de Biologie des Ligneux et des Grandes Cultures, UPRES EA 1207, UFR-Faculte des Sciences, Universite d'Orleans, rue de Chartres, BP 6759, 45067 Orleans cedex 2, France.[PMID: 15979315]

 Investigations have been made to develop an efficient protocol for micropropagation allowing to improve hypericin and pseudohypericin productions in Hypericum perforatum L. in vitro cultures. The role of growth regulator treatments has been particularly studied. Three in vitro culture lines with different morphological characteristics were obtained during H. perforatum micropropagation and referred to shoots, calli and plantlets according to their appearance. Multiplication and callogenesis from apical segments from sterile germinated seedlings were obtained on solid MS/B5 culture medium in the presence of N6-benzyladenine (BA) (0.1-5.0 mg/l BA). Regenerative potential of shoots was assessed on medium supplemented with auxins (0.05-1.0 mg/l), indole-3-acetic acid (IAA) or indole-3-butyric acid (IBA). The main goal of the research was to summarize the influence of plant growth regulators on hypericin and pseudohypericin productions in in vitro cultures of Hypericum. A rapid method for naphtodianthrone quantification was developed. The use of a reversed-phase high performance liquid chromatography (HPLC) method with fluorescence detection was used. Identification of the compounds was confirmed by electrospray ionization-mass spectrometry (ESI-MS) with electrospray in negative ion mode [M-H] . Calli, shoots and plantlets of H. perforatum produced hypericin and pseudohypericin. The concentration range of BA from 0.1 to 2.0 mg/l improved the production of hypericin (25-50 microg/g dry mass (DM)) and pseudohypericin (170-350 microg/g DM) in shoots. In callus cultures, BA (4.0-5.0 mg/l) did not changed hypericin contents (15-20 microg/g DM) but influenced pseudohypericin productions (120-180 microg/g DM). In the presence of auxins (IAA and IBA), Hypericum plantlets produced hypericin (30-100 microg/g DM) and pseudohypericin (120-400 microg/g DM). The presence of IAA did not influence naphtodianthrone productions in plantlets, but IBA decreased hypericin and pseudohypericin amounts in plantlets. The specific accumulation of the naphtodianthrones in in vitro cultures was influenced by phytohormonal supplementation of the medium. Results indicated that the production of hypericin and pseudohypericin could be increased by carefully adapted in vitro cultures. Hypericum in vitro cultures represent promising systems for hypericin and pseudohypericin productions.

   Reassessment of the toxicity of Hypericum perforatum (St John's wort) for cattle.:Aust Vet J. 2004 Nov;82(11):707-10.Bourke CA, White JG. NSW Agriculture, Orange Agricultural Institute, Forest Road, Orange, New South Wales.[PMID:15977618]

 OBJECTIVE: To investigate the clinical effect of administering sufficient Hypericum perforatum to cattle to deliver quadruple the reported oral toxic dose. ANIMALS: Thirty-six yearling Hereford (n = 18) and Angus (n = 18) steers. DESIGN: A series of six experiments was conducted, each using 12 animals in a 2 x 2 factorial design, with two breeds of cattle (Hereford, Angus) and two dose levels of hypericin, 1.5 mg/kg (treated group) and 0 mg/kg (control group). Each set of 12 steers was used in duplicate experiments, with all animals alternated between treated and control groups. PROCEDURES: Treated groups received finely milled H. perforatum administered orally in gelatin capsules to provide 1.5 mg hypericin/kg body weight. All cattle were then exposed to direct sunlight for 5 h per day for 5 successive days. Rectal temperatures were measured immediately before and at the end of each sunlight exposure session. Rectal temperature above 40 degrees C, together with some other clinical sign of hypericin poisoning, was considered indicative of intoxication. RESULTS: No animals developed a rectal temperature above 40 degrees C or other clinical signs of hypericin poisoning. CONCLUSIONS: While the reported bovine oral toxic dose of 3 g dried plant/kg body weight, for flowering stage, presumed narrow leaved biotype, H. perforatum, is probably correct, the corresponding dose for hypericin of 0.37 mg/kg is incorrect. Based on its known concentration in this plant the toxic dose of hypericin for partially pigmented Hereford-cross cattle is estimated at about 10.5 mg/kg body weight and more than this for fully pigmented cattle. This would imply that cattle of the former type should be about three and a half times better protected against H. perforatum toxicity than are unpigmented, wool protected, Merino sheep. Cattle, particularly if fully pigmented, may have a role in grazing management to control H. perforatum.

   Comparative chemical composition and variability of biological activity of methanolic extracts from Hypericum perforatum L.:Nat Prod Res. 2005 Apr;19(3):295-303.Conforti F, Statti GA, Tundis R, Bianchi A, Agrimonti C, Sacchetti G, Andreotti E, Menichini F, Poli F. Dipartimento di Scienze Farmaceutiche, Universita della Calabria, via Pietro Bucci, I-87036 Rende (CS), Italy.[PMID: 15702645]

 The biovariability of Hypericum perforatum L. (St. John's Wort) grown wild in Calabria and Sardinia (Italy) was reported with the aim to characterize the species through the isolation, detection, and quantitative evaluations of chemical markers (hypericin, quercetin, rutin) by HPLC analysis. Antioxidant activity of the methanolic H. perforatum extracts showed that the Calabrian samples were more active than those from Sardinia. The antibacterial activity evidenced the best performance on the gram positive bacteria with a MIC value of 50 microg/mL. Moreover, antifungal activity of all the extracts was also tested which showed interesting results particularly on the phytopathogene fungus P. ultimum. The variability shown by the samples could be attributed to environmental factors such as chemical-physical properties, composition of the soil, geographical coordinate, altitude, and solar exposure. The phytochemical analysis and the biological activity data suggested a possible use of H. perforatum extracts in the alimentary, cosmetic, and pharmaceutical fields.


   St John's wort (Hypericum perforatum) diminishes cognitive impairment caused by the chronic restraint stress in rats.:Pharmacol Res. 2005 Mar;51(3):239-46.Trofimiuk E, Walesiuk A, Braszko JJ. Department of Clinical Pharmacology, Medical University of Bialystok, Waszyngtona 15A, 15-274 Bialystok, Poland.[PMID: 15661574]

 In this study we tested the hypothesis that St John's wort (Hypericum perforatum) may counteract stress-induced memory impairment. Object recognition test and Morris water maze were used to determine whether administration of H. perforatum (350 mg kg(-1) for 21 days), standardized to 0.3% hypericin content, protects against non-spatial and/or spatial memory impairments due to chronic restraint stress (2h daily for 21 days). A group of rats administered the exogenous corticosterone at the dose of 5 mg kg(-1) daily for 21 days, yielding its similar plasma levels as these observed in stress was run in parallel. In the first experiment all rats were tested for recognition memory in the object recognition test. On the following day, the animals were tested in open field and elevated "plus" maze to control for the contribution of respectively, motor and emotional effects of our treatments to the memory tests. In the second experiment, new group of stressed animals was tested for spatial memory in the water maze. We observed that H. perforatum prevented the deleterious effects of both chronic restraint stress and long-term corticosterone on learning and memory as measured in both, the object recognition and the water maze tests. The herb not only prevented stress- and corticosterone-induced memory impairments, but it significantly improved recognition memory (p<0.01) in comparison to control. These results suggest that H. perforatum has a potential to prevent stress memory disorders.

   Effects of St John's wort and hypericin on cytotoxicity of anticancer drugs.:Drug Metab Pharmacokinet. 2002;17(5):467-74.

 St John's wort (SJW) is Hypericum perforatum L., Hypericaceae, a herbaceous perennial plant native to Europe and Asia, and its various preparations are widely used for the treatment of mild-to-moderately severe depressive disorders. With increasingly prevalent use, the interactions with SJW preparations with co-administered drugs have been reported, presumably via MDR1-mediated processes. In this paper, the effects of SJW extract on antiproliferative effects of anticancer drugs and the expression of MDR1 mRNA were examined using HeLa and its MDR1-overexpressing subline. The effects on MDR1-mediated transport were also evaluated using [(3)H]digoxin and LLC-GA5-COL150 cells, which were established by transfection of human MDR1 cDNA into porcine kidney epithelial LLC-PK(1) cells. The content of hypericin, a presumed active moiety within SJW extract, was determined by HPLC with a photo diode array to be 0.085 (w/w)%, and the effects of hypericin were also evaluated and compared with those of SJW extract. It was concluded that SJW extract reversed the cytotoxicity of paclitaxel and slightly of daunorubicin, down-regulated MDR1 mRNA, and inhibited MDR1-mediated transport, presumably due to other components than hypericin.

   Determining the quantity of hypericin in medicinal materials and asepsis seedings of Hypericum perforatum by HPLC.:Zhongguo Zhong Yao Za Zhi. 2003 Nov;28(11):1013-5.Chen ML, Huang LQ, Yang B, Fu MH, Luo YM. Institute of Chinese Materia Medica, Academy of Traditional Chinese Medicine, Beijing 100700.[PMID: 15615403]

 OBJECTIVE: To establish the method of determining the quantity of hypericin in Hypericum perforatum and determine the quantity of the hypericin in defferent medicinal materials and asepsis seedings which grow in defferent environment. METHOD: The specimen is extracted with methanol--Pyridine (9:1) ultrasound extraction. Chromatographic assay is performed on a hypersily ODS2 (4.6 mm x 150 mm, 5 microm) column. The mobile phase is composed of methanol -1.56% dihydric natrium phosphate hydrogen natrium solution (shift solution's acidity to 2.1 with phosphoric acid)--ethyl acetate (4:1.9:1), velocity of flow is 1 mL x min(-1); column temperature is 35 degrees C; the detection wavelength is 590 nm. RESULT: A satisfactory seperaration between hypericin and impurity. The calibration curve is linear over the range of 0.0524-0.2620 microg for hypericin (r = 0.9998). The average recovery of hypericin is 97.50%. CONCLUSION: The quantity of hypericin in Hypericum perforatum has something to do with the genetic factor, environment factor, growing period and dry means. The method of determining the quantity of hypericin can be regarded as the method of controling the quantity of medicinal materials.

   Influence of St John's wort on catecholamine turnover and cardiovascular regulation in humans.:Clin Pharmacol Ther. 2004 Nov;76(5):480-9.Schroeder C, Tank J, Goldstein DS, Stoeter M, Haertter S, Luft FC, Jordan J.Franz Volhard Clinical Research Center, Charite-Campus Buch and Helio Klinikum, Berlin, Germany.[PMID: 15536462]

 BACKGROUND: St John's wort (Hypericum perforatum) is a popular over-the-counter antidepressant. Its antidepressive effect has been attributed in part to inhibition of monoamine transporters and monoamine oxidase, on the basis of in vitro studies. METHODS: In a double-blind, randomized, placebo-controlled, crossover study, 16 healthy subjects (11 men and 5 women; mean age, 31 +/- 5 years) ingested either St John's wort (300 mg three times daily) or placebo for 7 days. Imipramine treatment (50 mg three times daily) in 7 subjects served as a positive control. After treatment, physiologic and biochemical tests included cardiovascular reflex testing, graded head-up tilt testing, and plasma catecholamine determinations. RESULTS: St John's wort had no effect on blood pressure, heart rate, heart rate variability, or blood pressure variability, regardless of the test condition. St John's wort had no effect on plasma concentrations of norepinephrine and its main metabolite, dihydroxyphenylglycol, whereas plasma dihydroxyphenylacetic acid (DOPAC; the main metabolite of dopamine) concentrations increased in every subject (1661 +/- 924 pg/mL versus 1110 +/- 322 pg/mL with placebo, P=.04). In contrast, imipramine increased resting blood pressure (124 +/- 10 mmHg/71 +/- 5 mmHg versus 110 +/- 8 mmHg/61 +/- 6 mmHg with placebo, P=.005 for systolic values and P=.003 for diastolic values) and heart rate (74 +/- 7 beats/min versus 62 +/- 6 beats/min with placebo, P=.005) and elicited a marked orthostatic tachycardia (increase in heart rate of 43 +/- 17 beats/min versus 26 +/- 8 beats/min with placebo, P=.006). CONCLUSIONS: Our findings challenge the concept that St John's wort elicits a major change in norepinephrine uptake or monoamine oxidase activity in vivo. The consistent increase in plasma DOPAC concentrations might suggest a novel mode of action or an inhibitory effect on dopamine beta-hydroxylase that should be followed up. We propose that a combination of physiologic and biochemical profiling may help better define the mode of action and potential side effects of herbal remedies.

   Quantitative analysis of active substances in St. John's wort (Hypericum perforatum L.) by the high performance liquid chromatography method:Medicina (Kaunas). 2004;40(10):975-81.Kazlauskas S, Bagdonaite E.Kaunas University of Medicine, A. Mickeviciaus 9, 44307 Kaunas, Lithuania. saulius_kazlauskas@gama.vtu.lt.[PMID: 15516821]

 The flavonoids (rutin, quercetin, and isoquercetin) and hypericin are the main active substances of St. John's wort (Hypericum perforatum). They support regeneration of human body cells, provide disinfection, and act as antidepressant, antiviral, and anti-inflammation tools. Such large spectrum of the applicable functions depends on the quantity of each substance. The objective of this paper is to determine the regularities of the quantitative variation of rutin, quercetin, isoquercetin and hypericin in flowers and leaves of St. John's wort in different habitats and different plant development phases. Plant material for the quantitative analysis was collected in 1998-1999. The high performance liquid chromatography (HPLC) was applied for the quantitative analysis of the active substances in St. John's wort. The flavonoids (rutin, quercetin, and isoquercetin) were identified under the UV waves of 254 nm, and hypericin - under the waves of 590 nm. Larger quantities of rutin were found in leaves, while those of quercetin, isoquercetin and hypericin - in flowers of St. John's wort. The highest accumulation of active substances was observed during the development of flowering buds and in flowering time. The quantitative variation of active substances in different habitats allows the selection of the best plant samples for their cultivation and conservation in field collections.


   Drug interactions with St John's wort : mechanisms and clinical implications.:Drug Saf. 2004;27(11):773-97.Mannel M.Ad libitum Medical Services, Berlin, Germany. mannel@ad-libitum-medical.com.[PMID: 15350151]

 The purpose of this paper is to review preclinical and clinical evidence relating to drug interactions with preparations of the medicinal herb St John's wort (Hypericum perforatum). A systematic literature search was carried out in three electronic databases up to June 2004. Information about case reports classified as St John's wort drug interactions was retrieved from the WHO Collaborating Centre for International Drug Monitoring and from the UK Medicines and Healthcare products Regulatory Agency in June 2003. Against the background of proven efficacy in mild to moderate depressive disorders and an excellent tolerability profile in monotherapy, there is sufficient evidence from interaction studies and case reports to suggest that St John's wort may induce the cytochrome P450 (CYP) 3A4 enzyme system and the P-glycoprotein drug transporter in a clinically relevant manner, thereby reducing efficacy of co-medications. Drugs most prominently affected and contraindicated for concomitant use with St John's wort are metabolised via both CYP3A4 and P-glycoprotein pathways, including HIV protease inhibitors, HIV non-nucleoside reverse transcriptase inhibitors (only CYP3A4), the immunosuppressants ciclosporin and tacrolimus, and the antineoplastic agents irinotecan and imatinib mesylate. Efficacy of hormonal contraceptives may be impaired as reflected by case reports of irregular bleedings and unwanted pregnancies. Drugs with a narrow therapeutic index should be monitored more closely when St John's wort is added, discontinued or the dosage is changed. The St John's wort constituent hyperforin is probably responsible for CYP3A4 induction via activation of a nuclear steroid/pregnane and xenobiotic receptor (SXR/PXR) and hypericin may be assumed to be the P-glycoprotein inducing compound, although the available evidence is less convincing. Combinations of St John's wort with serotonergic agents and other antidepressants should be restricted to prescription-only, by experienced clinicians, due to potential central pharmacodynamic interactions. In conclusion, providing certain precautions and contraindications are followed, and adequate information is given to healthcare professionals and patients, the safe and effective use of quality-tested St John's wort products can be ensured.

   CNS Spectr. 2001 Oct;6(10):835-40.Meltzer-Brody SE.Department of Psychiatry, University of Pittsburgh, Pittsburgh, PA, USA. Samantha_Meltzer@unc.edu .[PMID: 15334038].

 Depression and anxiety are among the top 10 health problems for which complementary and alternative therapies (CATs) are most frequently used, and medicinal herbs are among the most popular of these treatments. St. John's wort (Hypericum perforatum) is a perennial herb that has become a widely used depression therapy. Extracts of hypericum have shown affinity for receptors within multiple neurochemical systems. The primary active substance responsible for the antidepressant effect is not well defined, but most work has concentrated specifically on the hypericin and hyperforin components. Although hypericum has demonstrated significant antidepressant and antianxiety effects in multiple studies, there are several recent studies that do not support the previous evidence. In all reported studies, hypericum extracts have been well tolerated. In addition, new psychiatric uses for hypericum in obsessive-compulsive disorder, generalized anxiety disorder, menopausal symptoms, and alcohol dependence have been reported. Because patients are choosing to pursue CAT as a first-line therapy, psychiatrists will need to have a better understanding of phytomedicines used for treating depression and anxiety, and thus be better prepared to serve as effective allies of their patients.

   Neuroprotective effect of H. perforatum extracts on beta-amyloid-induced neurotoxicity.:Neurotox Res. 2004;6(2):119-30.Silva BA, Dias AC, Ferreres F, Malva JO, Oliveira CR. Institute of Biochemistry and Center for Neuroscience and Cell Biology of Coimbra, Faculty of Medicine, University of Coimbra, 3004-504 Coimbra, Portugal.[PMID: 15325964]

 In the present study we assessed the neuroprotective role of a Hypericum perforatum ethanolic extract and obtained fractions in amyloid-beta peptide (Abeta)((25-35))-induced cell death in rat cultured hippocampal neurons. Lipid peroxidation was used as a marker of oxidative stress by following the formation of TBARS in rat cortical synaptosomes, after incubation with ascorbate/Fe2+, alone or in the presence of EC97 effective concentrations of H. perforatum fractions. Induced lipid peroxidation was significantly inhibited by fractions containing flavonol glycosides, flavonol and biflavone aglycones, and by a fraction containing several phenols, mainly chlorogenic acid-type phenolics (21%, 77% and 98%, respectively). Lipid peroxidation evaluated after incubation with 25 microM Abeta(25-35), was significantly inhibited by H. perforatum extract. Cell viability was assessed by use of the Syto-13/PI assay. The total ethanolic extract (TE) and fractions containing flavonol glycosides, flavonol and biflavone aglycones, reduced Abeta(25-35)-induced cell death (65%, 58% and 59%, respectively). These results were further supported by morphological analysis of cells stained with cresyl violet. Peptide beta-amyloid(25-35) induced a decrease in cell volume, chromatin condensation and nuclear fragmentation, alterations not evident in the presence of the TE and fractions containing hypericins (hypericin concentration = 11.02 microM), or fractions containing flavonoids (quercetin concentration = 21.13 microM). Dendritic lesion, an evidence of neurodegeneration, was observed by neuronal staining with cobalt following insult with Abeta(25-35), but prevented after exposure to the peptide plus the fractions referred above. The results of the present paper suggest that H. perforatum extracts may be endowed with neuroprotective compounds able to prevent Abeta(25-35)-induced toxicity.

   The influence of extraction method on the quality of the liquid extract of St John's wort.:Medicina (Kaunas). 2004;40(8):745-9.Ramanauskiene K, Savickas A, Bernatoniene J. Department of Drug Technology and Pharmacy Organization, A. Mickeviciaus 9, 44307 Kaunas, Lithuania.[PMID: 15299991]

 The liquid extract of St John's wort was produced by methods of remaceration, percolation, and repercolation. Extraction by remaceration did not result in the complete extraction of herbal raw material; therefore, this method was rejected. Extraction by percolation requires the process of evaporation of the weak extract to take place, during which the active substances can decompose. This method was found to be not efficient. The liquid extract of St John's wort was produced by the complete-cycle repercolation: by dividing the raw material into unequal parts, and by dividing it into equal parts. It has been found that the highest concentration of hypericine, flavonoids and the dry remainder is obtained in the extract produced by repercolation with raw material divided into equal parts. The optimal method of extraction for St John's wort has been confirmed to be the complete-cycle repercolation with raw material divided into equal parts. In the course of the experiment, it was observed that the release of active substances from the raw material is influenced by the density of raw material filling. The optimal density of raw material filling has been found to be 0.11 g/cm3. The most suitable usage period was found to be 2 years.

   The antidepressant activity of Hypericum perforatum L. measured by two experimental methods on mice.:Acta Pharm. 2004 Jun;54(2):157-62.Bach-Rojecky L, Kalodjera Z, Samarzija I.Department of Pharmacology, Faculty of Pharmacy and Biochemistry, University of Zagreb, 10 000 Zagreb, Croatia. lbach@pharma.hr.[PMID: 15274759]

 The pharmacological approach to the treatment of depression includes a long-term employment of antidepressants, either in the form of monotherapy or as a combination of several antidepressants with various mechanisms of action. Hypericum perforatum L. (St. John's wort) is the only natural antidepressant. Several constituents of the extract, such as hypericin and hyperforin, seem to be important for this effect. H. perforatum is considered to be an effective alternative to other therapeutic agents in the treatment of mild to moderate depression. The paper describes the investigation of the antidepressant effect of H. perforatum (doses 7, 35 and 70 mg kg(-1) b. m.) on mice using the forced-swimming and tail-suspension methods. As an indicator of the antidepressant effect, it was shown that the immobility time of animals in the forced-swimming and tail-suspension experiments was shorter, i.e. the activity of the animals was higher. With single doses of extract suspension increasing from 7 over 35 to 70 mg kg(-1) the antidepressant effect increased in proportion by 10.1%, 25.8% and 38.6% in the swimming method, and by 12.7%, 16.5% and 24.5% in the tail-suspension method compared to controls. H. perforatum extract displays dose-dependent antidepressant effect at a dose as low as 7 mg kg(-1). Both models have proved to be equally valuable for demonstration of substances with a potential antidepressant effect.


   The antidepressant mechanism of Hypericum perforatum.:Life Sci. 2004 Jul 16;75(9):1021-7.Mennini T, Gobbi M. Istituto di Ricerche Farmacologiche Mario Negri, Via Eritrea 62, 20157, Milano, Italy. tiziana@marionegri.it.[PMID: 15207650]

 Clinical data indicate that hydroalcoholic extracts of Hypericum perforatum might be as valuable as conventional antidepressants in mild-to-moderate depression, with fewer side effects. One clinical trial using two extracts with different hyperforin contents indicated it as the main active principle responsible for the antidepressant activity. Behavioural models in rodents confirm the antidepressant-like effect of Hypericum extracts and also of pure hyperforin and hypericin. A hydroalcoholic extract lacking hyperforin also lacks the antidepressant-like effect. According to pharmacokinetic data and binding studies, it appears that the antidepressant effect of Hypericum extract is unlikely be due to an interaction of hypericin with central neurotransmitter receptors. The main in vitro effects of hyperforin (at concentrations of 0.1-1 microM) are non-specific presynaptic effects, resulting in the non-selective inhibition of the uptake of many neurotransmitters, and the interaction with dopamine D1 and opioid receptors. However, it is still not clear whether these mechanisms can be activated in vivo, since after administration of Hypericum extract brain concentrations of hyperforin are well below those active in vitro. In the rat, Hypericum extract might indirectly activate sigma receptors in vivo (through the formation of an unknown metabolite or production of an endogenous ligand), suggesting a new target for its antidepressant effects.

   How effective is St John's wort? The evidence revisited.:J Clin Psychiatry. 2004 May;65(5):611-7.Werneke U, Horn O, Taylor DM. Centre for the Economics in Mental Health, Institute of Psychiatry, London, United Kingdom. Ursula.Werneke@elcmht.nhs.uk.[PMID: 15163246]

 BACKGROUND: St. John's wort (Hypericum perforatum) has been identified as an effective treatment for depression in controlled studies and subsequent meta-analyses. However, 3 recently published large studies failed to demonstrate robust efficacy. Updated meta-analysis and assessment of publication bias may help determine the true effect of St. John's wort. METHOD: Meta-analysis to reevaluate the effectiveness of St. John's wort as an antidepressant, funnel plot analysis, and meta-regression to assess the impact of publication bias, small-study effects, and variation in trial characteristics were performed. We conducted 2 analyses: a reproduction of a recent meta-analysis including 15 studies (Meta-15) and a meta-analysis extended by the 3 studies published since then (Meta-18). The studies in Meta-15 were identified through MEDLINE and EMBASE searches conducted in June 2000. The search terms used were St. John's wort, hypericum, hypericin, depression, and antidepressant, and no language restrictions were applied. For both meta-analyses, we compared funnel plots, Begg's rank correlation, Egger's regression, trim and fill method, and meta-regression. RESULTS: In both analyses, effect sizes in recent studies were smaller than those reported in earlier studies; the addition of more recent studies into the analyses resulted in reduced effect size. In Meta-15, St. John's wort was significantly more effective than placebo with a risk ratio (RR) of 1.97 (CI = 1.54 to 2.53). In Meta-18, the RR was reduced to 1.73 (CI = 1.40 to 2.14). On funnel plot analysis, the Meta-18 plot proved to be much more skewed than the Meta-15 plot. Meta-regression showed that increase in effect size was associated with smaller sample size only. The impact of baseline severity of depression could not be evaluated as the studies used different versions of the Hamilton Rating Scale for Depression. CONCLUSION: St. John's wort may be less effective in the treatment of depression than previously assumed and may finally be shown to be ineffective if future trials confirm this trend.

   Quantitative characterization of direct P-glycoprotein inhibition by St John's wort constituents hypericin and hyperforin.:J Pharm Pharmacol. 2004 Jan;56(1):123-8.Wang EJ, Barecki-Roach M, Johnson WW.Drug Metabolism and Pharmacokinetics, Schering-Plough Research Institute, Lafayette, NJ 07848, USA. Erjia.wang@spcorp.com.[PMID: 14980009]

 The ATP-binding cassette transporter P-glycoprotein (P-gp) exerts a critical role in the systemic disposition of, and exposure to, lipophilic and amphipathic drugs, carcinogens, toxins and other xenobiotics. The ability of P-gp to transfer a wide variety of structurally unrelated compounds from the cell interior across the membrane bilayer remains intriguing. Since natural product chemicals in the widely consumed St John's wort appear to exert antidepressant effects by an unknown mechanism, the constituents are frequently studied for interactions with various biomacromolecules as well as cytotoxins or isolated cells. The drug interactions caused by this widely used herbal remedy are under-appreciated. Various clinical interactions have been observed upon the co-administration of St John's wort, and P-gp and CYP3A4 have been indicted as the cause. We characterized several St John's wort constituents for their interaction with P-gp and their specific effects on the P-gp export activity of several marker substrates. Two of these constituents, hyperforin and hypericin, inhibit the active efflux of the fluorescent markers daunorubicin (IC(50) approximately 30 microM) and calcein-AM. Herein, we show in-vitro results that can both explain the competing clinical observations of initial elevated exposure of P-gp substrate drugs (P-gp inhibition) followed by under-exposure (P-gp induction) when St John's wort is co-administered, and provide a further warning against unchecked co-administration of drugs with St John's wort.

   The effects of St. John's wort extract and amitriptyline on autonomic responses of blood vessels and sweat glands in healthy volunteers.:J Clin Psychopharmacol. 2004 Feb;24(1):79-82.[PMID: 14709951]Siepmann M, Kirch W, Krause S, Joraschky P, Mueck-Weymann M. Institute of Clinical Pharmacology, Medical Faculty, Technical University, Dresden, Germany. Martin.Siepmann@mailbox.tu-dresden.de

 St. John's wort extract is widely used and advertised as a "natural antidepressant" lacking autonomic side effects. This randomized, double-blind, placebo-controlled study compared the effects of St. John's wort extract on autonomic responses of blood vessels and sweat glands with those of amitriptyline and placebo. A randomized, double-blind, crossover study was performed in healthy male volunteers aged 22 to 31 years (25 +/- 3 years; mean +/- SD) years. Subjects orally received capsules with 255 to 285 mg St. John's wort extract (900 microg hypericin content), 25 mg amitriptyline, and placebo 3 times daily for periods of 14 days each with at least 14 days between. Vasoconstrictory response of cutaneous blood flow (VR) and skin conductance response (SR) following a single deep inspiration were employed as parameters of autonomic function. St. John's wort extract had no effect on VR and SR. In contrast, SR was diminished and the dilation phase of VR was prolonged following multiple dosing with amitriptyline (P < 0.05). Decreased electrodermal reactivity observed with amitriptyline reflects inhibition of acetylcholine at peripheral m3-cholinoreceptors, whereas prolongation of VR induced by the tricyclic drug may be due to sustained activation of central and/or peripheral sympathetic neurons.

   Extracts of St. John's wort and various constituents affect beta-adrenergic binding in rat frontal cortex.:Life Sci. 2004 Jan 9;74(8):1027-38.[PMID: 14672758]Simbrey K, Winterhoff H, Butterweck V.Institute of Pharmacology and Toxicology, Universitatsklinikum Munster, Domagkstrasse 12, 48149 Munster, Germany.

 The present study was designed to get further insight into the mode of antidepressant action of extracts prepared from St. John's wort (SJW) and relevant active constituents. Down-regulation of central beta-adrenergic receptors (beta-AR's) has been widely considered a common biochemical marker of antidepressant efficacy. Although previous studies have reported a beta-AR down-regulation for SJW extracts, in vivo studies that compare the effects of SJW extracts with those of relevant active constituents on beta-AR density have not been done yet. We used quantitative radioligand receptor-binding-studies to examine in rats the effects of short-term (2 wks) and long-term (8 wks) administration of different SJW extracts and constituents on beta-AR binding in rat frontal cortex. The effects were compared to those of the standard antidepressants imipramine and fluoxetine. [125I]CYP binding to beta-AR was found to be decreased after short as well as after long-term treatment with imipramine (36%, 40%). Short-term treatment with fluoxetine decreased the number of beta-adrenergic receptors (17%) while long-term treatment with fluoxetine elicited an increase (14%) in beta-AR-binding. This effect was comparable to that of the lipophilic CO2 extract which decreased beta-AR-binding (13%) after two weeks and slightly increased the number of beta-AR's after 8 weeks (9%). Short-term treatment with the methanolic SJW extract decreased beta-AR-binding (14%), no effects for this extract were observed after 8 weeks. Treatment with hypericin led to a significant down-regulation (13%) of beta-AR's in the frontal cortex after 8-weeks, but not after 2 weeks, while hyperforin (used as trimethoxybenzoate, TMB), and hyperoside were ineffective in both treatment paradigms. Compared to the SJW extracts and single compounds the effect of imipramine on beta-AR-binding was more pronounced in both treatment paradigms.


   Dissolution, solubility and cooperativity of phenolic compounds from Hypericum perforatum L. in aqueous systems.:Pharmazie. 2003 Mar;58(3):200-3.Jurgenliemk G, Nahrstedt A.Institut fur Pharmazeutische Biologie und Phytochemie, Westf. Wilhelms-Universitat, Munster, Germany.[PMID: 12685814]

 The dissolution in water of phenolic constituents of St. John's wort (Hypericum perforatum L.) from a medicinal tea and a coated tablet formulation showed different dissolution profiles. In general, the flavonoid glycosides were well dissolved, followed by flavonoid aglycones and hypericin while hyperforin was only detectable at a very low level. Interestingly, hypericin exhibited much better extraction and dissolution rates than the similarly lipophilic hyperforin. When determining the octanol/water partition coefficient it became obvious that the solubility of pure hypericin in water increased upon addition of some phenolic constituents typical for Hypericum extracts. Most effective in solubilizing hypericin was hyperoside (hyperin, quercetin 3-O-beta-D-galactoside) which increased the concentration of hypericin in the water phase up to 400 fold in this model.

   Hypericum perforatum, a source of neuroactive lead structures.:Curr Top Med Chem. 2003;3(2):187-201.Verotta L.Dipartimento di Chimica Organica e Industriale, Universita degli Studi di Milano, via Venezian 21, 20133 Milano, Italy. luisella.verotta@unimi.it.[PMID: 12570773]

 Although St. John's wort has been known for thousands of years and has been used for a variety of medicinal purposes, understanding of its activity and mechanisms of action is relatively new and not well understood. While researchers originally thought the naphthodianthrone hypericin was responsible for Hypericum's antidepressant activity, it is now believed some other compound or a combination of constituents exerts their antidepressant activity on the body. Hypericum is unique in that it seems to impact all known neurotransmitters at some level, directly, or indirectly through receptor sensitivity and regulation. There has been a proliferation of clinical studies on Hypericum in the last ten years, and even though some of these studies might be methodologically flawed, the preponderance of the evidence proves Hypericum to be beneficial for the treatment of mild-to-moderate depression, with a very favorable side effect profile. One clinical trial carried out using two extracts with different hyperforin content indicate this constituent as (one of) the main active principle responsible for the antidepressant activity.

   Long term administration of Hypericum perforatum improves spatial learning and memory in the water maze.:Biol Pharm Bull. 2002 Oct;25(10):1289-94.Widy-Tyszkiewicz E, Piechal A, Joniec I, Blecharz-Klin K.Department of Experimental and Clinical Pharmacology, Medical University of Warsaw, Poland. ewtpharm@amwaw.edu.pl.[PMID: 12392080]

 The aim of the present study is to investigate the effects of long-term Hypericum perforatum treatment on spatial learning and memory in rats. Hypericum preparation (HP) standardized to 0.3% hypericin content was administered orally for 9 weeks in doses of 4.3 and 13 microg/kg corresponding to therapeutic dosages in humans of 0.3 and 0.9 mg of total hypericins daily. A Morris water maze paradigm was used. The mean escape latency over 4 d for the Control group (21.9 s) and HP 4.3 group (21.7 s) was significantly greater than the latency of the HP 13 group (15.8s). In the probe trial on day 5, the HP 13 group crossed the correct annulus in the SE quadrant more often (4.5) than the other groups: Con (2.4) and HP 4.3 (3.1). After completion of the behavioral experiment, the regional brain concentrations of monoamines and metabolites were estimated in selected brain regions, i.e. prefrontal cortex, hippocampus and hypothalamus. Analysis of variance (ANOVA) demonstrated significant differences in the content of monoamines and metabolites between the treatment groups compared to the Control. The increased 5-hydroxytryptamine (5-HT) levels in the prefrontal cortex correlated positively with the retention of spatial memory. These findings show that the long-term administration of Hypericum perforatum can improve learning and spatial memory with significant changes in the content of monoamines in several brain regions.

   St. John's wort (Hypericum perforatum L.). A plant with relevance for dermatology.:Hautarzt. 2002 May;53(5):316-21.Schempp CM, Muller KA, Winghofer B, Schopf E, Simon JC. Universitats-Hautklinik, Haupstrasse, 7, 79104 Freiburg. schempp@haut.ukl.uni-freiburg.de.[PMID: 12063742]

 Saint John's wort (Hypericum perforatum L.) is a herbal remedy that is effective in the treatment of mild to moderate depression. In traditional folk medicine, oily extracts of St. John's wort are used for topical treatment of wounds, burns and myalgia. The lipophilic phloroglucin-derivative hyperforin has antibacterial and antiinflammatory effects. These effects could be of relevance in topical treatment of infected wounds and other dermatoses, but no studies have been conducted so far. The naphtodianthrone hypericin is a photodtodynamic active substance that kills tumor cells via the induction of apoptosis. Hypericin also displays antiviral activity in vitro. In vivo, intravenous or oral treatment with hypericin of HIV-infected subjects did not result in a reduction of the virus load. Most of the patients treated with hypericin experienced phototoxicity. Similar phototoxic symptoms ("hypericism") have been observed in grazing animals ingesting large amounts of St. John's wort. In contrast, antidepressant medication with St. John's wort usually does not produce phototoxic symptoms. Recent pharmacokinetic studies suggest that the phototoxic threshold level of hypericin is not reached with dosages used for the oral treatment of depression. However, very recent reports demonstrated interactions of St. John's wort with other drugs such as digoxin, indinavir and cyclosporin. Blood levels of these drugs were dramatically decreased by St. John's wort. This should be considered in the treatment of skin conditions with antiviral drugs or cyclosporin.

   St John's wort (Hypericum perforatum) modulates evoked potentials in guinea pig hippocampal slices via AMPA and GABA receptors.:Eur Neuropsychopharmacol. 2002 Jun;12(3):209-16.Langosch JM, Zhou XY, Heinen M, Kupferschmid S, Chatterjee SS, Noldner M, Walden J.Department of Psychiatry, University of Freiburg, Hauptstrasse 5, D-79104 Freiburg, Germany. jens_langosch@web.de.[PMID: 12007672]

 The effects of an ethanolic extract of the plant Hypericum perforatum L. (St John's wort) (HYP) and its hydrosoluble fraction (HYPWS) on electrically evoked population spikes and fEPSP were investigated in this study. Concentration dependent (10(-6) to 10(-4) g/l) excitatory effects were found. Above concentrations of 10(-3) g/l, HYP reduced the evoked responses, whereas HYPWS further increased them. Paired pulse facilitation was unaffected with HYPWS (10(-4) to 10(-2) g/l). The excitatory effects of HYPWS were amplified by the GABA(A) and GABA(B) receptor antagonists bicuculline and phaclofen, respectively. These excitations were antagonised by the AMPA receptor antagonist CNQX. Excitations caused by hypericum were not antagonised by the NMDA receptor antagonists D-APV and MK801, the metabotropic glutamate receptor (type I and II) antagonist MCPG, or the L-type calcium channel blocker verapamil. Hypericin and hyperforin, two components of H. perforatum, were found not to be responsible for the excitatory effects of the extract.


   St. John's wort (Hypericum perforatum)--is it safe during breastfeeding?.:Pharmacopsychiatry. 2002 Jan;35(1):29-30.Klier CM, Schafer MR, Schmid-Siegel B, Lenz G, Mannel M. Department of Psychiatry, General Hospital, University of Vienna, Wahringer Gurtel 18-20, 1090 Vienna, Austria. claudia.klier@univie.ac.at[PMID: 11819157]

 Both doctors and patients often treat postnatal depression with herbal preparations derived from St. John's wort. Because these preparations are available to patients as "natural" over-the-counter drugs for depression, they are popularly assumed to be safe. However, no systematic information exists regarding treatment of postnatal depression, infant's safety or pharmacokinetics of hypericum constituents in human breast milk or infant plasma. A mother with post-natal depression was admitted at our service. Her pharmacist had recommended taking a St. John's wort preparation three times a day (Jarsin 300, Lichtwer Pharma AG, Berlin, Germany). Four breast-milk samples (fore and hind milk) during an 18-hour period were analyzed to measure concentration of hypericin and hyperforin. Only hyperforin is excreted into breast milk at a low level, hyperforin and hypericin (two major active components) were below the lower limit of quantification (BLQ: below lower limit of quantification, LQ hypericin: 0.20 ng/ml, LQ hyperforin: 0.50 ng/ml) in this infant's plasma. No side effects were seen in the mother or infant. Before recommending St John's wort for the treatment of depression to women who breastfeed, long-term studies of outcome in infants are needed.

   Development of a simple, rapid and reproducible HPLC assay for the simultaneous determination of hypericins and stabilized hyperforin in commercial St. John's wort preparations.:J Pharm Biomed Anal. 2001 Dec;26(5-6):959-65.de los Reyes GC, Koda RT.Department of Pharmaceutical Sciences, University of Southern California, 1985 Zonal Ave., Los Angeles, CA 90089-9121, USA.[PMID: 11600308]

 A reversed-phase HPLC method was developed and validated for the simultaneous determination of hypericins and stabilized hyperforin in St. John's wort extract. The sample solution was prepared by extraction of the finely powdered extract with methanol-water (80:20, v/v) containing 5% HP-beta-cyclodextrin, and adjusted to pH 2.5 with orthophosphoric acid. Diluted extract solutions, maintained at 0 degrees C, were injected into a C18 column. The samples were eluted isocratically using a mobile phase consisting of acetonitrile and 0.3% v/v phosphoric acid (90:10, v/v) at a 1.5 ml/min flow rate with simultaneous fluorescence (315/590 nm, excitation/emission) and UV (273 nm) detection. Quantification of the marker compounds (hypericin, pseudohypericin, hyperforin) was achieved by use of standard curves generated by plotting peak heights versus concentrations. Validation studies demonstrated that this HPLC method is simple, rapid, reliable, and reproducible. The standard curves were linear over the concentration ranges, 0.5-2.5 microg/ml (hypericin), 0.35-1.6 microg/ml (pseudohypericin) and 5-50 microg/ml (hyperforin). The intra-day coefficients of variation obtained for hypericin, pseudohypericin and hyperforin were < or = 4.4%, < or = 5.4%, and < or = 2.8%, respectively; inter-day CVs were < or = 5.8%, < or = 4.9%, and < or = 2.5%, respectively. This method may be applied for the routine standardization of St. John's wort products against hyperforin and the hypericins, the putative antidepressant principles in the herbal.

   Pharmacological and endocrine effects of Hypericum perforatum and hypericin after repeated treatment.:Pharmacopsychiatry. 2001 Jul;34 Suppl 1:S2-7.Butterweck V, Korte B, Winterhoff H.Institute of Pharmacology and Toxicology, Munster, Germany. butterv@uni-muenster.de.[PMID: 11518068]

 Clinical studies have demonstrated the antidepressant efficacy of Hypericum (St. John's wort) extracts comparable to tricyclic antidepressants such as imipramine. We examined the effects of Hypericum extract and hypericin, one active constituent, in the forced swimming test (FST) after treatment repeated for 14 days. It has recently been shown that hypericin was inactive in the FST after acute treatment, but remarkably active when solubilized by subfraction IIIc1 containing mainly procyanidin B2. Therefore, we investigated the cooperative effects of hypericin and procyanidin B2 after repeated treatment. Imipramine (15 mg/kg), Hypericum extract (500 mg/kg) and hypericin (0.1 mg/kg) given daily for 2 weeks significantly reduced immobility time in the FST. No differences were observed between animals receiving pure hypericin and those receiving hypericin in combination with procyanidin B2. As several antidepressants act on the neuroendocrine axis resulting in altered hormone concentrations, selected endocrine parameters were investigated after repeated treatment. Daily treatment with either imipramine, Hypericum extract or hypericin alone or in combination with procyanidin B2 for 14 days significantly decreased plasma ACTH and corticosterone levels. None of the substances had pronounced effects on plasma prolactin or LH levels. From our present data, we propose that cooperative effects of hypericin and procyanidin B2 are of important relevance for the acute, but not for the chronic effects of this polycylic quinone.

   Neurochemical studies with St. John's wort in vitro.:Pharmacopsychiatry. 2001 Jul;34 Suppl 1:S137-42.Simmen U, Higelin J, Berger-Buter K, Schaffner W, Lundstrom K. Institute of Pharmaceutical Biology, University of Basel, Witterswil, Switzerland.[PMID: 11518063]

 The effect of extracts and constituents of St. John's wort, Hypericum perforatum, at various CNS receptors were studied by radioligand binding techniques in order to determine a profile of pharmacological activity in vitro. Binding inhibition was examined for the G-protein coupled opioid, serotonin (5-HT), histamine, neurokinin and corticotropin releasing factor (CRF) receptors, for the steroid estrogen-alpha receptor and for the ligand-gated ionchannel GABA(A) receptor. Hypericin showed the most potent binding inhibiton of all tested constituents to human CRF1 receptor with an IC50 value of 300 nM. Preliminary GTPgamma35S binding studies to CRF1 coupled G-protein indicated an antagonistic action for hypericin. The acylphloroglucinole hyperforin failed to inhibit 125I-astressin binding to hCRF, receptor up to 10 microM. Hyperforin inhibited binding to opioid and serotonin (5-HT) receptors at IC50 values between 0.4 and 3 microM, while hypericin and pseudohypericin inhibited with weaker potency. The biflavonoid I3,II8-biapigenin inhibited 3H-estradiol binding to the estrogen-alpha receptor with an IC50 value of 1 microM. The inhibition of 3H-muscimol binding to the GABA(A) receptor is likely to be exclusively due to GABA present in the extract. We therefore hypothesize that additive or synergistic actions of several ditsinct compounds may be responsible for the beneficial antidepressant effect of St. John's wort.

   Simultaneous determination of the predominant hyperforins and hypericins in St. John's Wort (Hypericum perforatum L.) by liquid chromatography.:J AOAC Int. 2000 Jul-Aug;83(4):944-9.Gray DE, Rottinghaus GE, Garrett HE, Pallardy SG, Gray DE, Rottinghaus GE, Garrett HE, Pallardy SG.University of Missouri, Department of Forestry, Columbia 65211, USA.[PMID: 10995119]

 Hypericin and hyperforin are believed to be among the active constituents in common St. John's wort (Hypericum perforatum L.). Presently, dietary supplements are generally standardized to contain specified levels of hypericin and hyperforin, and the related compounds, pseudohypericin and adhyperforin. A rapid method was developed for simultaneous determination of these 4 active constituents by liquid chromatography (LC). A 1 g portion of dried, finely ground leaf/flower sample is extracted with 20 mL methanol for 2 h. A 0.6 mL aliquot of the crude extract is combined with 5.4 mL acetonitrile-methanol (9 + 1) and passed through a mixed solid-phase cleanup column. The eluate is examined by LC for hyperforin, adhyperforin, hypericin, and pseudohypericin on a Hypersil reversed-phase column by using simultaneous ultraviolet (284 nm) and fluorescence detection (excitation, 470 nm; emission, 590 nm). The compounds are easily separated isocratically within 8 min with a mobile phase of acetonitrile-aqueous 0.1 M triethylammonium acetate (8 + 2). Average recoveries of hyperforin and adhyperforin were 101.9 and 98.4%, respectively, for 3 sample mixtures containing concentrations ranging from approximately 0.2 to 1.5% combined hyperforins per gram dry weight. Average relative standard deviation (RSD) values for hyperforin and adhyperforin for all 3 mixtures were 18.9 and 18.0%, respectively. Average recoveries of hypericin and pseudohypericin were 88.6 and 93.3% respectively, from 3 sample mixtures containing concentrations ranging from approximately 0.2 to 0.4% combined hypericins per gram dry weight. Average RSD values for hypericin and pseudohypericin for all 3 mixtures were 3.8 and 4.2%, respectively.


   Biochemical activities of extracts from Hypericum perforatum L. 1st Communication: inhibition of dopamine-beta-hydroxylase.:Arzneimittelforschung. 1999 Feb;49(2):106-9.Kleber E, Obry T, Hippeli S, Schneider W, Elstner EF.Lehrstuhl fur Phytopathologie, Labor fur angewandte Biochemie, Technische Universitat Munchen, Freising/Weihenstephan, Germany.[PMID: 10083977]

 Extracts from the herb "St. John's wort" (Hypericum perforatum L.) are used for the treatment of mental depression, nervousness, sleeplessness and for their wound healing, diuretic and antirheumatic properties. As one biochemical mechanism for depression lack of catecholamine neurotransmitters has been discussed. The results of this investigation show that alcoholic extracts from Hypericum perforatum L. on the basis of total hypericin content inhibit dopamine-beta-hydroxylase with an IC50 of 0.1 mu mol/l; pure commercial hypericin inhibits with an IC50 of 21 mu mol/l. Enzymes involved in the synthesis of dopamine from tyrosine, namely tyrosinase and tyrosine decarboxylase, are not influenced by hypericin at concentrations from 1 up to 10 mu mol/l.

   Biochemical activities of extracts from Hypericum perforatum L. 2nd Communication: inhibition of metenkephaline- and tyrosine-dimerization.:Arzneimittelforschung. 1999 Feb;49(2):109-14.Denke A, Schneider W, Elstner EF.Lehrstuhl fur Phytopathologie, Labor fur angewandte Biochemie, Technische Universitat Munchen, Freising-Weihenstephan, Germany.[PMID: 10083978]

 Extracts from the herb "St. John's wort" (Hypericum perforatum L.), besides other activities such as wound healing, antigout, antirheumatic and diuretic properties, are widely used to counteract neurological disorders such as depressive situations, nervousness and sleeplessness. The characteristic and leading component in these extracts, the dianthraquinone hypericin, is very likely not to represent the main active principle mediating the desirable effects. Thus, standardization of the drug is no longer based on the quantification of total hypericin and since several years simply the determination of dry matter content is in use instead. As biochemical background of depression the lack of catecholamine neurotransmitters or decreased beta-endorphins such as methionine- or leucine-enkephalins have to be envisaged. This communication reports on the inhibition of myeloperoxidase-catalyzed dimerization of enkephalins by Hypericum extracts. The substitution for enkephalins by tyrosine and for myeloperoxidase by horseradish peroxidase may represent a simple and inexpensive biochemical model reaction of pathological events during the manifestation of depressive events suitable for drug standardization.

   Biochemical activities of extracts from Hypericum perforatum L. 3rd Communication: modulation of peroxidase activity as a simple method for standardization.:Arzneimittelforschung. 1999 Feb;49(2):115-9.Schempp H, Denke A, Mann E, Schneider W, Elstner EF.Lehrstuhl fur Phytopathologie, Labor fur angewandte Biochemie, Technische Universitat Munchen, Freising-Weihenstephan, Germany.[PMID: 10083979]

 Alcoholic extracts from the herb "St. John's wort" (Hypericum perforatum L.) are widely used to counteract depressive situations, where the question on the mainly active principle is still under discussion. Thus, standardization of the drug on the basis of dry matter has been chosen instead of the popular leading component, hypericin. Inhibition of myeloperoxidase-catalyzed dimerization of enkephalins by Hypericum extracts has recently been reported. This method is based on the separation and quantification of enkephalin dimers by HPLC. In order to simplify this assay myeloperoxidase could be substituted by the cheaper horseradish peroxidase and the enkephalins by the amino acid tyrosine without loss of significance. In this communication we represent a more rapid photometric method based on peroxidase-catalyzed indole acetic acid oxidation suitable for quick, simple and economic drug standardization.

   Biochemical activities of extracts from Hypericum perforatum L. 4th Communication: influence of different cultivation methods.:Arzneimittelforschung. 1999 Feb;49(2):120-5.Denke A, Schempp H, Mann E, Schneider W, Elstner EF.Lehrstuhl fur Phytopathologie, Labor fur angewandte Biochemie, Technische Universitat Munchen, Freising-Weihenstephan, Germany.[PMID: 10083980]

 Extracts from the herb "St. John's wort" (Hypericum perforatum L.), know since ancient times as medical plant, besides other activities such as wound healing, antighout, antirheumatic and diuretic properties, is used in the treatment of mild to moderate depression. Increasing application continuously makes cultivation under controlled conditions of Hypericum perforatum L. more important. This report shows the results of three-years cultivation experiments with Hypericum perforatum L. The findings indicate that N-fertilizing yields more plant material but results in a drug with less quantity of secondary metabolites. Important components of the drug were quantified by HPLC and their activities were analyzed with the aid of biochemical test systems. Narrowleaved and broadleaved varieties of Hypericum perforatum L. show differences both in activity and quality. In this experiments the most active extract was a methanolic extract derived from non-fertilized, broadleaved plants.


  Scientific References:

  1.St. John's Wort Hypericum perforatum super function.
  2.Research update of St. John's Wort Hypericum perforatum and hypericin.