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Hypericum perforatum (HP) contains valuable and beneficial bioactive compounds that have been used to treat or prevent several illnesses. Encapsulation technology offers protection of the active compounds and facilitates to expose of the biologically active compounds in a controlled mechanism. Microcapsulation of the hydroalcoholic gum arabic and maltodextrin have hot been used as wall materials in the encapsulation of HP extract. Therefore, the optimum microencapsulation parameters of Hypericum perforatum (HP) hydroalcoholic extract were determined using response surface methodology (RSM) for the evaluation of HP extract. Three levels of three independent variables were screened using the one-way ANOVA. Five responses were monitored, including total phenolic content (TPC), 2,2-Diphenyl-1-picrylhydrazyl (DPPH), carr index (CI), hausner ratio (HR), and solubility. Optimum drying conditions for Hypericum perforatum microcapsules (HPMs) were determined: 180 °C for inlet air temperature, 1.04/1 for ratio of maltodextrin to gum arabic (w/w), and 1.98/1 for coating to core material ratio (w/w). TPC, antioxidant activity, CI, HR, and solubility values were specified as 316.531 (mg/g GAE), 81.912%, 6.074, 1.066, and 35.017%, respectively, under the optimized conditions. The major compounds of Hypericum perforatum (hypericin and pseudohypericin) extract were determined as 4.19 μg/g microcapsule and 15.09 μg/g microcapsule, respectively. Scanning electron microscope (SEM) analysis revealed that the mean particle diameter of the HPMs was 20.36 µm. Based on these results, microencapsulation of HPMs by spray drying is a viable technique which protects the bioactive compounds of HP leaves, facilitating its application in the pharmaceutical, cosmetic, and food industries. [ABSTRACT FROM AUTHOR]

St. John’s wort (Hypericum perforatum) is a medicinal plant with a rich history of traditional use. It has been shown to possess a range of beneficial health properties, including antioxidant and anti-inflammatory activities. In this study, the content of flavonoids and the antioxidant activity of commercially available dried and wild-grown samples were analyzed using the LC–MS/MS method. In addition, these samples were evaluated for their functional constituents, such as phenolic acids (ferulic, caffeic, chlorogenic, and gallic acids), quercetin, rutin, pseudohypericin, and hypericin using the liquid chromatography tandem mass spectrometry method. The most important antioxidant constituents in the samples analyzed were polyphenols with chlorogenic acid as the predominant compound. The content of the most important biocomponents with antidepressant activity was also analyzed. The results suggest that wild plants exposed to more stress factors have higher amount of compounds with antidepressant effects than plants grown in controlled conditions.

• Antioxidant activities of four Hypericum species have been evaulated. • Total phenolic contents and main flavonoids were determined. • H. perforatum extract had the highest hypericin and pseudohypericin amount. • All Hypericum extracts exhibited strong DPPH and total antioxidant activity. Total phenolic content, antioxidant activity, and the main constituents of three Hypericum species (H. perforatum, H. scabrum, and H. origanifolium) from Turkey had been investigated in this study. The quantification of main constituents (hypericin, and pseudohypericin) was performed by HPLC. The aerial parts of the plant extracts were screened in terms of their total phenolic content (TPC) and antioxidant activity tests including DPPH (2,2 diphenyl 1- picrylhdrazyl) radical scavenging activity, trolox equivalent antioxidant capacity (TEAC), ferric cyanide reducing (FRAP) antioxidant power assay, and total antioxidant activity by ferricthiocyanate (FTC). The highest TPC value (148.31 ± 4.57 mg GAE/g DW) was obtained for H. scabrum (HS) while H. perforatum (HP) extract had the highest hypericin (9.57 ± 0.07 µg/mL), and pseudohypericin (7.82 ± 0.05 µg/mL) amount. All Hypericum species demonstrated stronger DPPH activities than the standard compounds butylated hydroxytoluene (BHT) and ascorbic acid (AA) with the values of IC 50 < 3.8 µg/mL. The highest trolox equivalent antioxidant capacity (TEAC) value (11.28 ± 0.28) was achieved with HO. Considerable values were obtained for HS (90.25 ± 0.05), HP (90.20 ± 0.07), and HO (88.42 ± 0.02) by total antioxidant determination using ferricthiocyanate (FTC) method with 2 days incubation. This study reveals that all Hypericum species are good sources of natural antioxidants with high TPC and major constituent contents. [ABSTRACT FROM AUTHOR]

For almost two years, the COVID-19 pandemic has constituted a major challenge to human health, particularly due to the lack of efficient antivirals to be used against the virus during routine treatment interventions. Multiple treatment options have been investigated for their potential inhibitory effect on SARS-CoV-2. Natural products, such as plant extracts, may be a promising option, as they have shown an antiviral activity against other viruses in the past. Here, a quantified extract of Hypericum perforatum was tested and found to possess a potent antiviral activity against SARS-CoV-2. The antiviral potency of the extract could be attributed to the naphtodianthrones hypericin and pseudohypericin, in contrast to other tested ingredients of the plant material, which did not show any antiviral activity. Hypericum perforatum and its main active ingredient hypericin were also effective against different SARS-CoV-2 variants (Alpha, Beta, Delta, and Omicron). Concerning its mechanism of action, evidence was obtained that Hypericum perforatum and hypericin may hold a direct virus-blocking effect against SARS-CoV-2 virus particles. Taken together, the presented data clearly emphasize the promising antiviral activity of Hypericum perforatum and its active ingredients against SARS-CoV-2 infections. [ABSTRACT FROM AUTHOR]

Chen XG, Hua F, Wang SG, Xu YY, Yue HT, Sun J. Drug Des Devel Ther. 2018;12:2389–2402The authors have advised Figures 1 (on page 2393), Figure 3 (on page 2394), Figure 5 (on page 2395), Figure 7 and Figure 8 (on page 2396) are incorrect due to a fault in their microscope camera. The correct Figures are shown below:Read the original article

Xiao-Gang Chen,1 Fu Hua,2 Shou-Guo Wang,1 Yong-Yi Xu,1 Hai-Tao Yue,1 Jin Sun1 1Department of Orthopedics, Huai’an First People’s Hospital, Nanjing Medical University, Huai’an, Jiangsu 223300, People’s Republic of China; 2Department of Gynaecology, Huai’an First People’s Hospital, Nanjing Medical University, Huai’an, Jiangsu 223300, People’s Republic of China Background: Biosynthesis of leukotriene (LT) by arachidonic acid involves 5-lipoxygenase (5-LO) as an important precursor. Here, we evaluated the role of pseudohypericin (PHP) for its postulated 5-LO inhibitory activity along with a Cys-LT receptor antagonist zafirlukast (ZFL) against inflammatory response and tissue injury in mice. Materials and methods: The spinal injury was induced by two-level laminectomy of T6 and T7 vertebrae. The inflammation was assessed by histology, inflammatory mediators by enzyme-linked immunosorbent assay, apoptosis by Annexin-V, FAS staining, terminal deoxynucleotidyltransferase-mediated UTP end labeling (TUNEL) assay and expression of Bax and Bcl-2 by Western blot. Effect on motor recovery of hind limbs was evaluated for 10 days postinjury. Results: The spinal injury resulted in tissue damage, apoptosis, edema, infiltration of neutrophils with increased expression of tumor necrosis factor-α (TNF-α) and cyclooxygenase-2 (COX-2). The spinal tissue showed elevated levels of prostaglandin E2 (PGE2), and LTB4 and increased phosphorylation of injured extracellular signal-regulated kinase-1/2 (ERK1/2). The PHP, ZFL and combination decreased inflammation, tissue injury and infiltration of neutrophils. Treatment also decreased the levels of PGE2, phosphorylation of extracellular signal-regulated kinase-1/2 (pERK 1/2), LT, TNF-α and COX-2 with a marked reduction in apoptosis and improved the motor function. Conclusion: The present study confirmed 5-LO antagonist activity of PHP and established its neuroprotective role along with ZFL. Keywords: pseudohypericin, zafirlukast, 5-lipoxygenase, Cys-LT, mice

Herbal medicines are frequently used for the prevention and treatment of obesity and obesity-related disorders. Our preliminary screening showed that St. John's Wort (SJW) displayed potent inhibition on pancreatic lipase (PL), a key hydrolase responsible for lipid digestion and absorption in mammals. Herein, the inhibition potentials and inhibitory mechanism of SJW extract and its major constituents on PL were fully investigated by a set of in vitro and in silico studies. The results clearly demonstrated that the naphthodianthrones, biflavones and most of flavonoids in SJW displayed strong to moderate inhibition on PL. Among all tested natural compounds, two naphthodianthrones (hypericin and pseudohypericin) and one biflavone (I3,II8-biapigenin) isolated from SJW exhibited potent PL inhibition activity, with the IC 50 values of <1 μM. Inhibition kinetics analyses showed that hypericin, pseudohypericin and I3,II8-biapigenin inhibited PL via a mixed manner, while molecular dynamics simulations revealed that three newly identified PL inhibitors could bind on PL at both the catalytic cavity and the interface between colipase and the C-terminal domain of PL. Collectively, our findings suggested that part of major constituents in SJW displayed potent PL inhibition activities, which could be used as lead compounds for the development of novel PL inhibitors. • The inhibitory effects of St. John's wort extract and its major constituents on pancreatic lipase (PL) are assayed. • Hypericin, pseudohypericin and I3,II8-biapigenin display potent inhibition on PL, with the IC 50 values of <1 μM. • Hypericin, pseudohypericin and I3,II8-biapigenin are mixed inhibitors of PL. • Hypericin, pseudohypericin and I3,II8-biapigeninc could bind on PL at two distinct ligand-binding sites. [ABSTRACT FROM AUTHOR]

Background/Aims: Pseudohypericin (P-HY) and its congener hypericin are the major hydroxylated phenanthroperylenediones present in Hypericum species. Our previous study indicated that hypericin was able to function as a sonosensitizer. The potential use of P-HY as a sonosensitizer for sonodynamic therapy (SDT) requires further exploration. Thus, this study aimed to investigate the effects of P-HY-SDT on THP-1 macrophages. Methods: THP-1 macrophages were incubated with P-HY, and cell viability was measured using a CCK-8 assay. Fluorescence microscopy assessed the intracellular reactive oxygen species (ROS), mitochondrial membrane potential (∆Ψm) and mitochondrial permeability transition pore (mPTP) opening. Apoptotic and necrotic cell levels were measured by the flow cytometry analysis. Western blots were employed to assay BAX, Cytochrome C expression and apoptosis-related proteins. Results: P-HY-SDT induced THP-1 macrophage apoptosis. The levels of ROS were significantly increased in the SDT group, resulting in both mPTP opening and ∆Ψm loss, which led to apoptosis. In addition, the translocation of BAX, release of Cytochrome C and the upregulated expression of apoptosis-related proteins after P-HY-SDT were observed, all of which were reversed by N-acetyl cysteine (NAC). Conclusion: P-HY-SDT induced THP-1 macrophage apoptosis through the mitochondria-caspase pathway via ROS generation, the translocation of BAX and the release of Cytochrome C to regulate the mPTP opening.

Background/Aims: Pseudohypericin (P-HY) and its congener hypericin are the major hydroxylated phenanthroperylenediones present in Hypericum species. Our previous study indicated that hypericin was able to function as a sonosensitizer. The potential use of P-HY as a sonosensitizer for sonodynamic therapy (SDT) requires further exploration. Thus, this study aimed to investigate the effects of P-HY-SDT on THP-1 macrophages. Methods: THP-1 macrophages were incubated with P-HY, and cell viability was measured using a CCK-8 assay. Fluorescence microscopy assessed the intracellular reactive oxygen species (ROS), mitochondrial membrane potential (∆Ψm) and mitochondrial permeability transition pore (mPTP) opening. Apoptotic and necrotic cell levels were measured by the flow cytometry analysis. Western blots were employed to assay BAX, Cytochrome C expression and apoptosis-related proteins. Results: P-HY-SDT induced THP-1 macrophage apoptosis. The levels of ROS were significantly increased in the SDT group, resulting in both mPTP opening and ∆Ψm loss, which led to apoptosis. In addition, the translocation of BAX, release of Cytochrome C and the upregulated expression of apoptosis-related proteins after P-HY-SDT were observed, all of which were reversed by N-acetyl cysteine (NAC). Conclusion: P-HY-SDT induced THP-1 macrophage apoptosis through the mitochondria-caspase pathway via ROS generation, the translocation of BAX and the release of Cytochrome C to regulate the mPTP opening. © 2016 The Author(s) Published by S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

The influence of the main parameters on extraction of naphthodianthrone pigments (hypericin and pseudohypericin) from raw phytomass Hypericum perforatum using microwave activation was studied. The maximal pigment extraction was found to be achieved when using 55% ethanol or isopropanol as extractants at a water duty of 40 (power density of microwave irradiation was 0.0205 W/cm at a frequency 2450 MHz for 60 s). The microwave-assisted extraction of hypericin and pseudohypericin from H. perforatum was experimentally proved to be a particular example of the mass transfer intensification in a capillary porous body-liquid system. Microwave activation is found to implement extraction by ten times, reducing the time necessary to fully extract the pigments from H. perforatum compared to classical extraction. [ABSTRACT FROM AUTHOR]

St. John's wort has attracted particular attention because of its beneficial effects as an antidepressant, antiviral, and anticancer agent. A method for the combination of integrated expanded bed adsorption chromatography and countercurrent chromatography for the simultaneous extraction and purification of pseudohypericin and hypericin from the herb is presented in this paper. Firstly, the constituents were extracted and directly adsorbed by expanded bed adsorption chromatography under optimal conditions. The stepwise elution was then performed by expanded bed adsorption chromatography that enriched the targets with higher purities and recoveries compared to other methods. Secondly, the eluent fractions from expanded bed adsorption chromatography were further separated by two-step high-speed countercurrent chromatography. A two-step high-speed countercurrent chromatography method with a biphasic solvent system composed of n-hexane/ethyl acetate/methanol/water with a volume ratio of 1:2:1:2 was performed by stepwise changing the flow rate of the mobile phase. Consequently, 5.6 mg of pseudohypericin and 2.2 mg of hypericin with purities of 95.5 and 95.0%, respectively, were successfully obtained from 40 mg of crude sample. [ABSTRACT FROM AUTHOR]

The molecularly imprinted polymers (MIPs) were prepared by an oxidation-reduction polymerization system using a non-covalent molecularly imprinting strategy with hypericin as the template, acrylamide as the functional monomer and pentaerythritol triacrylate as the cross-linker in the porogen of acetone. The UV spectrum revealed that a cooperative hydrogen-bonding complex between hypericin and acrylamide might be formed at the ratio of 1:6 in the prepolymerized system. Two classes of the binding sites were produced in the resulting hypericin-imprinted polymer with the dissociation constants of 16.61 μg L-1 and 69.35 μg L-1, and the affinity binding sites of 456.53 μg g-1 and 603.06 μg g-1, respectively. The synthesized MIPs were characterized by scanning electron microscope, thermogravimetric and differential thermal analysis. High-performance liquid chromatography was used to investigate the adsorption and recognition properties of the MIPs. Selective binding of the template molecule was demonstrated in comparison to the analog pseudohypericin. After the Hypericum perforatum L. plant being air dried and finely ground, an extract was prepared by shaking the powder in a methanol-water solution (80:20, v/v), vacuum filtration though a Büchner funnel, liquid-liquid extraction with ethyl ether and ethyl acetate, and evaporating on a rotary evaporator until dry. With the sorbents of the optimized MIPs, a molecularly imprinted solid-phase extraction (MISPE) procedure was developed for enrichment and separation of hypericin from the Hypericum extract in the presence of interfering substances. The selective extraction of hypericin from herbal medicine was achieved with the recovery of 82.30%. The results showed that MISPE can be a useful tool for specific isolation and effective clean-up of target compounds from natural products. [ABSTRACT FROM AUTHOR]

Hypericum perforatum(Hp) extracts contain many different classes of constituents including flavonoids and biflavonoids, phloroglucinols, naphthodianthrones, caffeic acid derivatives, and unknown and/or unidentified compounds. Many constituents may be responsible for the anti-inflammatory activity of Hp including quercetin and derivatives, hyperforin, pseudohypericin, and amentoflavone. In line with antidepressant data, it appears that the interactions of constituents may be important for the anti-inflammatory activity of Hp. Interactions of constituents, tested in bioavailability models, may explain why synergistic mechanisms have been found to be important for antidepressant and antiproliferative bioactivities. This review highlights the relationship among individual constituents and the anti-inflammatory activity of Hp extracts and proposes that interactions of constituents may be important for the anti-inflammatory activity of botanical extracts, although the exact mechanisms of the interactions are still unclear. [ABSTRACT FROM PUBLISHER]

This study reports the first quantification study of pseudohypericin (PsHyp) and hypericin (Hyp) inHypericum undulatumSchousb. ex Willdin vitrocultures developed by a Portuguese company. Both compounds were quantified by high-performance liquid chromatography and their levels were compared with those in commercial samples ofHypericum perforatum. PsHyp was found to be the major naphthodianthrone ofH. undulatum,with an average ratio of 3.73:1 compared to Hyp. Significant statistical differences were found between the content of Hyp and PsHyp inH. undulatumregenerated shoots compared toin vivosamples. The mean concentration of total Hyps varied from 178.41 to 358.93 µg g−1dry extract inH. undulatumregenerated shoots, which is on average two to three times less than naphthodianthrone levels found inH. undulatum in vivoandH. perforatumcommercial samples. However, none of the analysed samples presented the levels of Hyps required by the European and United States Pharmacopoeias. [ABSTRACT FROM PUBLISHER]

Hypericum perforatum is a well known medicinal plant. The main pharmacological properties are due to the presence of naphtodianthrones such as hypericin and pseudohypericin. Unfortunately the levels of these compounds vary under different environmental conditions. Elicitation of in vitro cultures is a useful approach to enhance and extend production of desirable products. Therefore, the effects of salicylic acid were characterized on different explants of H. perforatum L. (cells, calli and shoots) cultured in vitro. It appears at first that salicylic acid did not affect growth and development of these explants. In addition, the production of both hypericin and pseudohypericin has doubled in elicited cell suspension cultures but not in the two other cultures. Furthermore, phenylpropanoids that are among the most frequently observed metabolites affected upon treatment of in vitro culture material with elicitors, were produced and the enzymatic activities of phenylalanine ammonia lyase and of chalcone isomerase were stimulated upon elicitation. These effects were dependant of the type of in vitro culture, the concentration of salicylic acid and the duration post-elicitation. The H. perforatum cells were globally more sensitive to salicylic acid elicitation when maintained in an undifferentiated state and particularly in cell suspension cultures. In the absence of glands considered as the sites of naphtodianthrones biosynthesis, cells and calli were capable of producing these compounds. This implies that salicylic acid could act at biosynthesis level but not for the accumulation of both hypericin and pseudohypericin. Consequently, the regulation of this process is more complex than cited in the literature involving the responsibility of only Hyp- 1 gene, encoding a hypericin biosynthetic enzyme, cloned and characterized from H. perforatum. [ABSTRACT FROM AUTHOR]

Abstract: Hypericum perforatum (St. John’s wort) is an herb widely used as supplement for mild to moderate depression. Our prior studies established synergistic anti-inflammatory activity associated with 4 bioactive compounds in a fraction of a H. perforatum ethanol extract. Whether these 4 compounds also contributed to the ethanol extract activity was addressed in the research reported here. Despite the popularity of H. perforatum, other Hypericum species with different phytochemical profiles could have their anti-inflammatory potentials attributed to these or other compounds. In the current study, ethanol extracts of different Hypericum species were compared for their inhibitory effect on LPS-induced prostaglandin E2 (PGE2) and nitric oxide (NO) production in RAW 264.7 mouse macrophages. Among these extracts, those made from H. perforatum and H. gentianoides demonstrated stronger overall efficacy. LC–MS analysis established the 4 compounds were present in the H. perforatum extract and pseudohypericin in all active fractions. The 4 compounds accounted for a significant part of the extract’s inhibitory activity on PGE2, NO, tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) in RAW 264.7 as well as peritoneal macrophages. Pseudohypericin was the most important contributor of the anti-inflammatory potential among the 4 compounds. The lipophilic fractions of H. gentianoides extract, which did not contain the previously identified active constituents, decreased PGE2 and NO potently. These fractions were rich in acylphloroglucinols, including uliginosin A that accounted for a proportion of the anti-inflammatory activity observed with the active fractions. Overall, the current study established that a different group of major anti-inflammatory constituents were present in H. gentianoides, while showing that the previously identified 4 compound combination was important for H. perforatum’s anti-inflammatory potential. [Copyright &y& Elsevier]

We investigated the effects of plant growth regulators [6-benzyladenine (BA), kinetin (Kin), 6-γ,γ-dimethylallylaminopurine (2iP), thidiazuron (TDZ) and α-naphthaleneacetic acid (NAA)], modified Murashige and Skoog (MS) medium containing 10 mM NH and 5 mM NO and supplemented with 2iP, BA, Kin and NAA (MSM medium), and two elicitors [jasmonic acid (JA), and salicylic acid (SA)], on plant growth and accumulation of hypericins (hypericin and pseudohypericin) and hyperforin in shoot cultures of Hypericum hirsutum and H. maculatum. Our data suggested that culture of shoots on MS medium supplemented with BA (0.4 mg l) or Kin (0.4 mg l) enhanced production of hypericins in H. maculatum and hyperforin in H. hirsutum. Hypericins and hyperforin concentrations decreased in both species when TDZ (0.4 mg l) was added to the MS medium. Also, TDZ induced hyperhydric malformations and necrosis of regenerated shoots. Cultivation of H. maculatum on MSM medium resulted in approximately twofold increased production of hypericins compared to controls, and the growth of H. hirsutum shoots on the same medium led to a 6.16-fold increase in hyperforin production. Of the two elicitors, SA was more effective in stimulating the accumulation of hypericins. At 50 μM, SA enhanced the production of hypericin (7.98-fold) and pseudohypericin (13.58-fold) in H. hirsutum, and, at 200 μM, enhanced the production of hypericin (2.2-fold) and pseudohypericin (3.94-fold) in H. maculatum. [ABSTRACT FROM AUTHOR]

Abstract: Hypericin and hypericin-like substances are considered the main active compounds in Hypericum perforatum L. (Hypericaceae). In this work pseudohypericin and hypericin of H. perforatum collected in Lithuania were quantified. Studies on accumulation dynamics and between-accession variation of the contents of these secondary metabolites were carried out by high performance liquid chromatography (HPLC). The data were statistically processed with ANOVA and PCA. Significant difference between pseudohypericin and hypericin content in floral budding and full flowering stages was detected. The highest amounts of the secondary metabolites were observed in the flowering stage. The study revealed evident within population variations in H. perforatum. Mean concentrations of pseudohypericin and hypericin among accessions varied from 3.45 to 6.82 mg/g and from 1.17 to 2.59 mg/g, respectively. Accessions of H. perforatum showed remarkable differences in chemical composition depending on the provenance of plants. [ABSTRACT FROM AUTHOR]

Abstract: Hypericum perforatum extracts have been used to treat diseases, including mild-to-moderate depression and inflammatory conditions. It is particularly important to identify which constituents present in the H. perforatum extracts are responsible for its anti-inflammatory activity since consumers are taking H. perforatum preparations to treat inflammation. We used a combination of four putative bioactive constituents, called the 4-component-system that interacted synergistically to explain the light-activated anti-inflammatory activity of an H. perforatum fraction in RAW 264.7 mouse macrophages. We also combined the constituents at concentrations detected in the fraction to identify key molecular targets. LPS was used to model an inflammatory response, and the 4-component-system and H. perforatum fraction were used as treatments that inhibited LPS-induced prostaglandin E2 (PGE2) production in RAW 264.7 mouse macrophages in the studies of gene expression profiles. We used Affymetrix genechips, statistical analysis, and quantitative real-time PCR to identify key gene targets of the 4-component-system and the sub-fraction from an H. perforatum ethanol extract. The H. perforatum sub-fraction, with or without LPS stimulation, affected far more genes than the 4-component-system with and without LPS. Genes involved in Janus kinase, as well as a signal transducer and activator of transcription (JAK–STAT) and eicosanoid pathways were identified that could account for the reduction in PGE2 observed with both treatments in LPS-stimulated macrophages. Ten genes may be particularly important targets for activity of the 4-component-system and the fraction with LPS stimulation and these genes were involved in inflammatory signaling pathways, namely the JAK–STAT and eicosanoid pathways. [Copyright &y& Elsevier]

70 years have passed since the first isolation of the naphthodianthrones hypericin and pseudohypericin from Hypericum perforatum L. Today, they continue to be one of the most promising group of polyphenols, as they fascinate with their physical, chemical and important biological properties which derive from their unique chemical structure. Hypericins and their derivatives have been extensively studied mainly for their antitumor, antiviral and antidepressant properties. Notably, hypericin is one of the most potent naturally occurring photodynamic agents. It is able to generate the superoxide anion and a high quantum yield of singlet oxygen that are considered to be primarily responsible for its biological effects. The prooxidant photodynamic properties of hypericin have been exploited for the photodynamic therapy of cancer (PDT), as hypericin, in combination with light, very effectively induces apoptosis and/or necrosis of cancer cells. The mechanism by which these activities are expressed continues to be a main topic of discussion, but according to scientific data, different modes of action (generation of ROS & singlet oxygen species, antiangiogenesis, immune responces) and multiple molecular pathways (intrinsic/extrinsic apoptotic pathway, ERK inhibition) possibly interrelating are implicated. The aim of this review is to analyse the most recent advances (from 2005 and thereof) in the chemistry and biological activities (in vitro and in vivo) of the pure naphthodianthrones, hypericin and pseudohypericin from H. perforatum. Extracts from H. perforatum were not considered, nor pharmakokinetic or clinical data. Computerised literature searches were performed using the Medline (PubMed), ChemSciFinder and Scirus Library databases. No language restrictions were imposed. [ABSTRACT FROM AUTHOR]

The genus Hypericum (Guttiferae) is a source of biologically active secondary metabolites, notably hyperforin, hypericins and various phenolics. In the present study the presence of the phloroglucinol derivative hyperforin, the naphthodianthrones hypericin and pseudohypericin, the phenylpropane chlorogenic acid and the flavonoids such as rutin, hyperoside, apigenin-7- O-glucoside, kaempferol, quercitrin, quercetin and amentoflavone were investigated in two Turkish species of Hypericum, namely, Hypericum scabrum L. and Hypericum bupleuroides Gris. The aerial parts representing a total of 30 individuals were collected at full flowering and dissected into floral, leaf and stem tissues. After being dried at room temperature, the plant materials were assayed for secondary metabolite concentrations by HPLC. All of the secondary metabolites examined were detected in both species at various levels depending on plant tissue except for hyperforin which was not accumulated in Hypericum scabrum. The presence of hyperforin and the phenolic compounds examined in both species were reported by us for the first time. [ABSTRACT FROM AUTHOR]

The principle medicinal secondary metabolites present in Hypericum species are thought to be naphthodianthrones hypericin and pseudohypericin. The present study was conducted to determine ontogenetic and morphogenetic variations of hypericin and pseudohypericin contents in Hypericum triquetrifolium growing in two sites located in Northern Turkey. Plants were harvested at vegetative, flowering and fructification stages. Plants were dissected into stem, leaf and reproductive tissues at the time of harvest, dried separately and assayed for hypericin and pseudohypericin contents by high-performance liquid chromatography. The hypericin and pseudohypericin concentrations in the leaves and reproductive parts were highest at flowering and fructification, whereas the stem contents of these compounds decreased with advancing plant growth in both sites. The leaves contained higher concentrations of both naphthodianthrones than the stems or reproductive parts of H. triquetrifolium. [ABSTRACT FROM AUTHOR]

Abstract: Hypericum perforatum (Hp) has been used medicinally to treat a variety of conditions including mild-to-moderate depression. Recently, several anti-inflammatory activities of Hp have been reported. An ethanol extract of Hp was fractionated with the guidance of an anti-inflammatory bioassay (lipopolysaccharide (LPS)-induced prostaglandin E2 production (PGE2)), and four constituents were identified. When combined together at concentrations detected in the Hp fraction to make a 4 component system, these constituents (0.1μM chlorogenic acid (compound 1), 0.08μM amentoflavone (compound 2), 0.07μM quercetin (compound 3), and 0.03μM pseudohypericin (compound 4)) explained the majority of the activity of the fraction when activated by light, but only partially explained the activity of this Hp fraction in dark conditions. One of the constituents, light-activated pseudohypericin, was necessary, but not sufficient to explain the reduction in LPS-induced PGE2 of the 4 component system. The Hp fraction and the 4 component system inhibited lipoxygenase and cytosolic phospholipase A2, two enzymes in the PGE2-mediated inflammatory response. The 4 component system inhibited the production of the pro-inflammatory cytokine tumor necrosis factor-α (TNF-α), and the Hp fraction inhibited the anti-inflammatory cytokine interleukin-10 (IL-10). Thus, the Hp fraction and selected constituents from this fraction showed evidence of blocking pro-inflammatory mediators but not enhancing inflammation-suppressing mediators. [Copyright &y& Elsevier]

Abstract: The genus Hypericum has received considerable interest from scientists, as it contains the variety of structurally diverse natural products which possess a wide array of biological properties, mainly hypericins and hyperforin. In the present study, variations of pseudohypericin and hyperforin were investigated in two Turkish species of Hypericum, namely Hypericum perfoliatum and Hypericum origanifolium. Wild growing plants were harvested at vegetative, floral budding, flowering, fresh fruiting and mature fruiting stages, and dissected into stem, leaf and reproductive tissues and assayed for chemical contents by high performance liquid chromatography method. Content of pseudohypericin and hyperforin in samples of the whole plant increased during the course of ontogenesis in both species. The highest levels of the chemicals were reached at full flowering (2.62mg/g dry weight (DW) pseudohypericin and 1.84mg/gDW hyperforin for H. perfoliatum; 0.93mg/gDW pseudohypericin and 1.63mg/gDW hyperforin for H. origanifolium). Among different reproductive parts, full opened flowers produced the highest amount of pseudohypericin (1.18mg/gDW) and hyperforin (4.36mg/gDW) in H. origanifolium. Similarly, the highest pseudohypericin accumulation was observed in full opened flowers in H. perfoliatum (7.41mg/gDW) while floral buds of this species produced the highest amount of hyperforin (7.80mg/gDW). These data can be useful when elucidating the medicinal properties of the species and the chemosystematic significance of hyperforin and pseudohypericin in the relationships among species of Hypericum. [Copyright &y& Elsevier]

Hypericum perforatum is a perennial medicinal plant known as “St. John's wort” in Western Europe and has been used in the treatment of several diseases for centuries. In the present study, morphologic, phenologic and population variability in pseudohypericin and hyperforin concentrations among H. perforatum populations from Northern Turkey was investigated for the first time. The aerial parts of H. perforatum plants representing a total of 30 individuals were collected at full flowering from 10 sites of Northern Turkey to search the regional variation in the secondary metabolite concentrations. For morphologic and phenologic sampling, plants from one site were gathered in five phenological stages: vegetative, floral budding, full flowering, fresh fruiting and mature fruiting. The plant materials were air-dried at room temperature and subsequently assayed for chemical concentrations by high performance liquid chromatography. Secondary metabolite concentrations ranged from traces to 2.94 mg/g dry weight (DW) for pseudohypericin and traces –6.29 mg/g DW for hyperforin. The differences in the secondary metabolite concentrations among populations of H. perforatum were found to be significant. The populations varied greatly in hyperforin concentrations, whereas they produced a similar amount of pseudohypericin. Concentrations of both secondary metabolites in all tissues increased with advancing of plant development and higher accumulation levels were reached at flowering. Among different tissues, full opened flowers were found to be superior to stems, leaves and the other reproductive parts with regard to pseudohypericin and hyperforin accumulations. The present findings might be useful to optimize the processing methodology of wild-harvested plant material and obtain increased concentrations of these secondary metabolites. [ABSTRACT FROM AUTHOR]

Hypericin and pseudohypericin content in stems, leaves, and flowers of some Hypericum species growing in Turkey, namely, H. heterophyllum Vent, H. hyssopifolium L., H. linarioides Bosse, H. monbretii Spach, H. orientale L., H. origanifolium Willd., H. perforatum L., H. scabrum L., and H. triquetrifolium Turra, was determined by HPLC. Hypericin and pseudohypericin were detected in all species tested except for H. heterophyllum, and the presence in H. orientale and H. scabrum was reported by us for the first time in the current study. Hypericin and pseudohypericin contents observed in the study varied greatly depending on species and plant tissues. The lowest levels of hypericin and pseudohypericin were detected in leaves of H. hyssopifolium [0.030 and 0.051 mg/g dry weight (DW), respectively] whereas flowers of H. montbretii produced the highest levels of both hypericin forms (2.52 mg/g DW hypericin and 3.58 mg/g DW pseudohypericin). H. montbretti and H. triquetrifolium were found to be superior over H. perforatum with regard to hypericin and pseudohypericin content, encouraging the cultivation and biological evaluation of these species in Turkey. [ABSTRACT FROM AUTHOR]

Abstract: The 1H NMR spectra of the commercially available compounds hypericin and its derivative pseudohypericin in CD3OH solutions indicate significantly deshielded signals in the region of 14–15ppm. These resonances are attributed to the peri hydroxyl protons OH(6), OH(8) and OH(1), OH(13) of hypericins which participate in a strong six-membered ring intramolecular hydrogen bond with CO(7) and CO(14), respectively, and therefore, they are strongly deshielded. In the present work, we demonstrate that one-dimensional 1H NMR spectra of hypericin and pseudohypericin, in Hypericum perforatum extracts show important differences in the chemical shifts of the hydroxyl groups with excellent resolution in the region of 14–15ppm. The facile identification and quantification of hypericin and its derivative compound pseudohypericin was achieved, without prior HPLC separation, for two H. perforatum extracts from Greek cultivars and two commercial extracts: a dietary supplement, and an antidepressant medicine. The results were compared with those obtained from UV–vis and LC/MS measurements. [Copyright &y& Elsevier]

The present study was conducted to determine the content of pharmacologically important constituents hypericin, pseudohypericin, chlorogenic acid, rutin, hyperoside, apigenin-7-O-glucoside, quercitrin, quercetin and vitexin in eight Hypericum species namely, H. aviculariifolium Jaup. and Spach subsp. depilatum (Freyn and Bornm.) Robson var. depilatum (endemic), H. lydium Boiss., H. montbretii Spach, H. orientale L, H. origanifolium Willd, H. perfoliatum L, H. perforatum L. and H. pruinatum Boiss. and Bal. growing in different locations of Turkey. Wild growing plants were harvested at flowering and after dried subsequently assayed for the constituents by HPLC method. Hyperoside, quercitrin and pseudohypericin were found in all species. Hypericin, quercetin and chlorogenic acid were also detected in all species with the exceptions of H. orientale for hypericin, H. montbretii for quercetin and H. lydium for chlorogenic acid. Apigenin-7-O-glucoside accumulation was observed in all examined species at various levels, except for H. orientale and H. origanifolium. Rutin was detected in H. aviculariifolium, H. lydium, H. orientale, H. perfoliatum and H. perforatum. On the contrary of the other species, vitexin was found in only H. motbretii. The presence of flavonoid vitexin in the genus Hypericum was reported for the first time. [ABSTRACT FROM AUTHOR]

Abstract: Water stress is known to increase the secondary metabolites concentration in plant tissues and severe water stress conditions may cause oxidative stress due to the formation of reactive oxygen species and photoinhibitory damage. Current study was undertaken to evaluate the changes in the physiological status especially the photosynthetic efficiency and the biochemical profile of the leaf tissues of St. John''s wort (Hypericum perforatum) plants exposed to water stress. The net photosynthetic rates of the leaves of plants grown under a water stress condition were significantly low compared with those of the control. The maximal quantum efficiency of PSII photochemistry () of the dark adopted leaves was similar for both wilted and recovered plants although these values were significantly low compared with those of the control. The leaf tissues of the plants under a water stress condition had a significantly higher capacity to detoxify oxygen radicals with an about 2.5-fold increase over the antioxidant potential of the leaves of non-treated (control) or recovered plants. In the leaf tissues of plants grown under a water stress condition, both hypericin and pseudohypericin concentrations reduced with time and on Day 12 of the treatment, the concentration was significantly lower than that of the control; in contrast, the hyperforin concentration increased significantly and the value was nearly double after 12 days of the treatment. Under a water stress condition, the hyperforin concentration was about 70-fold higher than the total hypericins concentration in the leaf tissues; in case of control it was only 10–12 times higher. The results also indicate that the major secondary metabolite, hyperforin concentration of 62-day-old plants was three- to four-fold higher than the previously reported values for 1–2-year-old field-grown plants. [Copyright &y& Elsevier]

Phytochemical profiles and DNA fingerprints were obtained from 11 species and cultivars of Hypericum (H. androsaemum, H. calycinum, H. frondosum var. "Sunbrust", H. grandiflorum, H. inodorum, H. moseranum, H. olympicum, H. patulum vat. "Sungold", H. perforatum, H. pelforatum var. "Anthos", and H. perforatum vat. "Topas"). Accessions were identified using amplified fragment length polymorphism (AFLP) and DNA fingerprints. Although each accession had a distinctive DNA fingerprint, correlations between structurally similar plants and their DNA profiles were apparent. DNA banding patterns between each accession genetic distances were analyzed and quantified using a dendogram software program. High performance liquid chromatography (HPLC) was used to quantify the content of chlorogenic acid, hypertbrin, hypericin, pseudohypericin, and rutin in methanolic extracts of each Hypericum accession. Hypericum perforatum accessions and H. inodorum contained the highest concentration of marker phytochemicals, while H. calycimon, H. grandiflorum, and H. olympicunt contained the lowest concentration. AFLP profiles of the plants were correlated with their levels of marker phytochemicals enabling true to type identification and marker-assisted breeding programs. [ABSTRACT FROM AUTHOR]

Abstract: The polar methanolic fraction (PMF) of the Hypericum perforatum L. extract has recently been developed and tested as a novel, natural photosensitizer for use in the photodynamic therapy (PDT), and photodynamic diagnosis (PDD). PMF has been tested on HL-60 leukemic cells and cord blood hemopoietic progenitors. In the present study, the efficacy of PMF as a phototoxic agent against urinary bladder carcinoma has been studied using the T24 (high grade metastatic cancer), and RT4 (primary low grade papillary transitional cell carcinoma) human bladder cancer cells. Following cell culture incubation, PMF was excited using 630nm laser light. The photosensitizer exhibited significant photocytotoxicity in both cell lines at a concentration of 60μg/ml, with 4–8J/cm2 light dose, resulting in cell destruction from 80% to 86%. At the concentration of 20μg/ml PMF was not active in either cell line. These results were compared with the results obtained in the same cell lines, under the same conditions with a clinically approved photosensitizer, Photofrin®. Photofrin® was used in the maximum clinically tolerable dose of 4μg/ml, and it was also excited with 630nm laser light. In the T24 cell Photofrin® exhibited slightly less photocytotocixity, compared with PMF, resulting in 77% cell death with 8J/cm2 light dose. However, against the RT4 cells Photofrin® resulted in minimal cell death (9%) with even 8J/cm2 light dose. Finally, the type of cell death induced by PMF photoactivation was studied using flow cytometry and DNA laddering. Cell death by PMF photodynamic action in these two bladder cell lines is caused predominently by apoptosis. The reported significant photocytotoxicity, selective localization, natural abundance, easy, and inexpensive preparation, underscore that the PMF extract hold the promise of being a novel, effective PDT photosensitizer. [Copyright &y& Elsevier]

Abstract: 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 590nm 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. [Copyright &y& Elsevier]

Abstract: 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. [Copyright &y& Elsevier]

Abstract: St. John''s wort (Hypericum perforatum L.) plants grown in a field or a greenhouse have been used for the preparation of phytopharmaceutical products. However, fields and greenhouse have such changeable environments, which can lead to high variations in the concentrations of bioactive compounds. In addition, due to the changeable environment, studies aiming to determine the effect of a unique environmental factor, the harvesting time or the germplasm on concentrations of bioactive compounds cannot be conclusive. In the current study, plants were grown in a growth chamber under controlled environments, and the effect of two different temperatures, 25 and 30°C, was determined on concentrations of the major bioactive compounds. Moreover, the variation in the concentrations of the major bioactive compounds with harvesting time, and germplasm lines were investigated. The concentrations of major bioactive compounds in the shoots reached a peak on day 52, regardless of the temperature. Throughout the experiment, pseudohypericin and hypericin concentrations were higher at 30°C than at 25°C, whereas hyperforin concentrations remained higher at 25°C than at 30°C. Moreover, high variations in the concentrations of the major bioactive compounds were found among the different germplasm lines. [Copyright &y& Elsevier]

Abstract: The analysis of hypericin, pseudohypericin (collectively called in this study hypericins) and hyperforin in biological fluids is reported using single-drop liquid-phase microextraction in conjunction with HPLC-UV-fluorescence detection. A new option for analysis of the active principle constituents in biological samples is proposed, reducing the steps required prior to analysis. There are several parameters which determine the mass transfer such as the extraction solvent, drop and sample volumes, extraction time and temperature, pH and ionic strength, stirring rate and depth of needle tip in the bulk solution. These parameters were chosen to optimize the performance in the current study. The method was validated with respect to precision, accuracy and specificity. The intra-day precision values were below 2.3% for the high concentration level of control samples and 6.2% for the low level. The respective inter-day precision values were calculated to be below 4.4 and 7.1%, respectively, for the two concentration levels. Accuracy of the method, calculated as relative error, ranged from −2.6 to 7.0%. It was demonstrated that as long as the extraction procedure is consistently applied, quantitative analysis is performed accurately and reproducibly in human urine and plasma samples. Limits of quantitation (LOQs) in urine were calculated to be 3, 6 and 12ng/ml for pseudohypericin, hypericin and hyperforin, respectively. Slightly higher limits were measured in plasma, i.e. 5, 12 and 20ng/ml, for the respective analytes. [Copyright &y& Elsevier]

Abstract: Temperature stress is known to cause many physiological, biochemical and molecular changes in plant metabolism and possibly alter the secondary metabolite production in plants. The hypothesis of the current study was that temperature stress can increase the secondary metabolite concentrations in St. John''s wort. Plants were grown under controlled environments with artificial light using cool white fluorescent lamps and CO2 enrichment and 70-day-old plants were subjected for 15 days to different temperature treatments of 15, 20, 25, 30 and 35 °C before harvested. Major aim of the study was to increase the major secondary metabolites in St. John''s wort by applying temperature stress and to evaluate the physiological status of the plant especially the photosynthetic efficiency and peroxidase activity of the leaf tissues exposed to different temperatures under precisely controlled environmental factors. Results revealed that relatively high (35 °C) or low (15 °C) temperatures reduced the photosynthetic efficiency of the leaves of St. John''s wort plants and resulted in low CO2 assimilation. Net photosynthetic rates and the maximal quantum efficiency of PSII photochemistry of the dark adopted leaves (ϕp max) decreased significantly in the leaves of plants grown under 35 or 15 °C temperature treatments. High temperature (35 °C) treatment increased the leaf total peroxidase activity and also increased the hypericin, pseudohypericin and hyperforin concentrations in the shoot tissues. These results provide the first indication that temperature is an important environmental factor to optimize the secondary metabolite production in St. John''s wort and controlled environment technology can allow the precise application of such specific stresses. [Copyright &y& Elsevier]

Copyright of Drug Research / Arzneimittel-Forschung (Editio Cantor Verlag fur Medizin und Naturwissenschaften) is the property of Editio Cantor Verlag fur Medizin und Naturwissenschaften and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Abstract: Hypericum perforatum L. (St. John''s wort), a traditional herb with antidepressive and wound healing properties, was grown under 100, 300 and 600μmolm−2 s−1 photosynthetic photon flux (PPF) each with 500, 1000 and 1500μmolmol−1 CO2, with the objective of maximizing the biomass and secondary metabolite production. Plants grown in the field (1770μmolm−2 s−1 PPF and 380μmolmol−1 CO2) were used as a control. On day 45, total fresh and dry mass, and the number of stem nodes were greatest in plants grown under 600μmolm−2 s−1 PPF with 1500μmolmol−1 CO2 concentration (HH-treatment) and were 29, 30 and 4 times greater, respectively, than those of the control. Leaf net photosynthetic rate (Pn) increased with increasing PPF and/or CO2 concentration, with the highest value occurring in the plants grown under the HH-treatment. Secondary metabolite contents (mg/plant) in the leaf tissues were generally higher in the plants grown under the controlled environments than those in the field (control). Hypericin and pseudohypericin contents were highest in the HH-treatment, being 30 and 41 times greater, respectively, than those of the control. Hypericin and pseudohypericin contents in the leaf tissues increased with increasing Pn. The second order polynomial correlations between Pn and hypericin, and between Pn and pseudohypericin contents with R 2 of 0.82 and 0.79, respectively, were obtained. Moreover, total hypericin (hypericin+pseudohypericin) concentration (mgg−1 leaf DM) in leaf tissues was related to both PPF and CO2 concentration as expressed by second order polynomial correlations (R 2 =1). Therefore, growing St. John''s wort plants under a controlled environment can enhance biomass and secondary metabolite production by increasing net photosynthetic rate. [Copyright &y& Elsevier]

Abstract: 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 μg/g dry mass (DM)) and pseudohypericin (170–350 μg/g DM) in shoots. In callus cultures, BA (4.0–5.0 mg/l) did not changed hypericin contents (15–20 μg/g DM) but influenced pseudohypericin productions (120–180 μg/g DM). In the presence of auxins (IAA and IBA), Hypericum plantlets produced hypericin (30–100 μg/g DM) and pseudohypericin (120–400 μg/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. [Copyright &y& Elsevier]

Hypericum perforatum L. (St. John’s wort) is a traditional medicinal plant that has been used for the treatment of neurological disorders and depression. To investigate a large-scale in vitro growth system for St. John’s wort cv ‘New Stem’, six different culture systems, balloon type bubble bioreactor, temporary immersion bioreactor, temporary root zone immersion bioreactor, Erlenmeyer flask, large vessel with gelled medium under forced ventilation (LFV system), and Magenta vessel, were compared with greenhouse-grown plants for biomass production efficiency and accumulation of the selected bioactive molecules, hypericin, pseudohypericin, and hyperforin. After 25 days in culture, significantly more plant fresh mass was observed in a balloon type bubble (BB) bioreactor system than in other in vitro culture systems or in the greenhouse system. Plant dry mass was at a similar level in BB bioreactor, LFV system, and greenhouse. Chlorophylls a and b were highest in the leaves of plantlets grown in either large vessels with forced ventilation or temporary root zone immersion bioreactors. In general, levels of hypericin, pseudohypericin and hyperforin of the plantlets grown in the LFV system and levels of hypericin and pseudohypericin in the Magenta vessel were greater or similar to those in greenhouse-grown plants. Gelled medium (LFV system and Magenta vessel) had higher concentrations of hypericin and pseudohypericin than those grown in liquid medium. Together these results show that St. John’s wort can be efficiently produced in a variety of in vitro culture systems for aseptic production of bioactive compounds. [Copyright &y& Elsevier]

The ability for synthesis of hypericins and phloroglucinols was studied in in vitro grown Hypericum perforatum seedlings. HPLC analysis of hypericin, pseudohypericin and their immediate precursors protohypericin and protopseudohypericin, and phloroglucinols, hyperforin and adhyperforin, revealed great intraspecific variation. A significant correlation was found between the total content of hypericins and phloroglucinols and between the content of hypericin/protohypericin and pseudohypericin/protopseudohypericin. The presence of hyperforin and adhyperforin was first proved in the Hypericum perforatum seedlings at early stage of development. Peculiarities in reproduction pathways do not primarily affect the formation of secondary metabolites but significantly contribute to genetic variation. Genetic factors as ploidy level, inheritance of the mode of reproduction, coexistence of several reproductive pathways in a particular plant that contribute to chemical variation, which is expressed in the early stages of ontogeny are discussed. [Copyright &y& Elsevier]

At high pH, pseudohypericin from St. John''s Wort (Hypericum perforatum) is converted into a single product which has been isolated. LC–MS analysis indicated that the molar mass of this product is identical to that of pseudohypericin. 1H and 13C NMR analysis indicated that the compound is a novel phenanthroperylene quinone which we have named isopseudohypericin; it is a cyclic ether which is an isomer of pseudohypericin. [Copyright &y& Elsevier]

St. John''s wort (Hypericum perforatum L.) is widely used for the treatment of mild to moderately severe depression. However, the nature of its active principles and the exact mode of antidepressant action are still unknown. It has been suggested repeatedly in preclinical and clinical studies that the content of the acylphloroglucinol hyperforin decisively contributes to the antidepressant efficacy of St. John''s wort extracts. Experimental studies in vivo also indicate that the naphthodianthrone hypericin may reduce the activity of the hypothalamic–pituitary–adrenal axis. Exacerbated hypothalamic–pituitary–adrenal activity has often been associated with depressive states in patients. Corticotropin-releasing factor (CRF) seems to be a major determinant in the regulation of the hypothalamic–pituitary–adrenal activity via activation of CRF1 receptors. In the present study, we investigated the CRF1 receptor antagonist activity of three main constituents of St. John''s wort (hypericin, pseudohypericin and hyperforin) by measuring their effect on CRF-stimulated cAMP formation in recombinant Chinese hamster ovary (CHO) cells. As a selectivity test, the compounds were also tested against calcitonin in the same cells. Of the three compounds tested, only pseudohypericin selectively antagonised CRF (KB 0.76 μM). Hypericin and hyperforin affected both CRF and calcitonin with similar potencies and the same type of behaviour (competitive antagonism for hypericin, noncompetitive for hyperforin). It is concluded that pseudohypericin is the only real CRF1 receptor antagonist of the three constituents tested. In addition, evidence is provided that beside hyperforin, both pseudohypericin and hypericin are implicated in the antidepressant efficacy of St. John''s wort. [Copyright &y& Elsevier]

The effect of increased levels of carbon dioxide on the growth (fresh weight), morphogenesis (formation of leaves, roots and shoots) and tissue concentrations of the naphthodianthrones (hypericin and pseudohypericin) was determined for St. John's Wort (Hypericum perforatum L.). Plants, started from shoot cuttings in a vermiculite-peat moss mixture within a greenhouse employing natural sunlight, were grown for eight weeks under CO2 levels of 350, 1500, 3000, 10,000 and 30,000 µl CO2/liter atmospheres. Elevated CO2 levels (≥ 1500 µl CO2/liter) significantly increased growth and morphogenesis compared with ambient (350 µl CO2/liter) levels (control). Levels of hypericin were significantly higher at 1500 and 3000 µl CO2/liter than at ambient CO2, while the higher (10,000 and 30,000 µl 2/liter) CO2 atmospheres caused reductions in levels of both naphthodianthrones in plant tissues. Because of increased plant growth, however, naphthodianthrone yields were significantly increased at elevated CO2 levels as compared with controls. [ABSTRACT FROM PUBLISHER]

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The stimulating effect of cork pieces on hypericin and pseudohypericin biosyntheses was studied in cells of shoots regenerated from the callus cultures of St. John's wort ( Hypericum perforatumL.). The addition of the cork matrix slightly stimulated shoot growth and enhanced pseudohypericin biosynthesis about threefold (to 0.4 mg/g dry wt). Pseudohypericin production increased proportionally with the amount of cork material added (from 1 to 4 mg/ml of growth medium). Further increase in the amount of cork pieces inhibited both pseudohypericin production and shoot growth. Organic and aqueous extracts of cork pieces did not affect the production of these substances. [ABSTRACT FROM AUTHOR]