Hyperforin Content Research Articles

St. John's wort extract with a high hyperforin content does not induce P-glycoprotein activity at the human blood-brain barrier.

St. John's wort (SJW) extract, a herbal medicine with antidepressant effects, is a potent inducer of intestinal and/or hepatic cytochrome P450 (CYP) enzymes and P-glycoprotein (P-gp), which can cause clinically relevant drug interactions. It is currently not known whether SJW can also induce P-gp activity at the human blood-brain barrier (BBB), which may potentially lead to decreased brain exposure and efficacy of certain central nervous system (CNS)-targeted P-gp substrate drugs. In this study, we used a combination of positron emission tomography (PET) imaging and cocktail phenotyping to gain a comprehensive picture on the effect of SJW on central and peripheral P-gp and CYP activities. Before and after treatment of healthy volunteers (n = 10) with SJW extract with a high hyperforin content (3-6%) for 12-19 days (1800 mg/day), the activity of P-gp at the BBB was assessed by means of PET imaging with the P-gp substrate [11C]metoclopramide and the activity of peripheral P-gp and CYPs was assessed by administering a low-dose phenotyping cocktail (caffeine, omeprazole, dextromethorphan, and midazolam or fexofenadine). SJW significantly increased peripheral P-gp, CYP3A, and CYP2C19 activity. Conversely, no significant changes in the peripheral metabolism, brain distribution, and P-gp-mediated efflux of [11C]metoclopramide across the BBB were observed following the treatment with SJW extract. Our data suggest that SJW does not lead to significant P-gp induction at the human BBB despite its ability to induce peripheral P-gp and CYPs. Simultaneous intake of SJW with CNS-targeted P-gp substrate drugs is not expected to lead to P-gp-mediated drug interactions at the BBB.

Apolar Extracts of St. John's Wort Alleviate the Effects of β-Amyloid Toxicity in Early Alzheimer's Disease.

Hypericum perforatum (St. John's wort) has been described to be beneficial for the treatment of Alzheimer's disease (AD). Different extractions have demonstrated efficiency in mice and humans, esp. extracts with a low hypericin and hyperforin content to reduce side effects such as phototoxicity. In order to systematically elucidate the therapeutic effects of H. perforatum extracts with different polarities, APP-transgenic mice were treated with a total ethanol extract (TE), a polar extract obtained from TE, and an apolar supercritical CO2 (scCO2) extract. The scCO2 extract was formulated with silicon dioxide (SiO2) for better oral application. APP-transgenic mice were treated with several extracts (total, polar, apolar) at different concentrations. We established an early treatment paradigm from the age of 40 days until the age of 80 days, starting before the onset of cerebral β-amyloid (Aβ) deposition at 45 days of age. Their effects on intracerebral soluble and insoluble Aβ were analyzed using biochemical analyses. Our study confirms that the scCO2H. perforatum formulation shows better biological activity against Aβ-related pathological effects than the TE or polar extracts. Clinically, the treatment resulted in a dose-dependent improvement in food intake with augmentation of the body weight, and, biochemically, it resulted in a significant reduction in both soluble and insoluble Aβ (-27% and -25%, respectively). We therefore recommend apolar H. perforatum extracts for the early oral treatment of patients with mild cognitive impairment or early AD.

St. John's wort formulations induce rat CYP3A23-3A1 independent of their hyperforin content.

The pregnane X receptor (PXR) is a ligand activated regulator of cytochrome P450 (CYP)3A enzymes. Among the ligands of human PXR is hyperforin, a constituent of St John's wort (SJW) extracts and potent inducer of human CYP3A4. It was the aim of this study to compare the effect of hyperforin and SJW formulations controlled for its content on CYP3A23-3A1 in rats. Hyperiplant® was used as it contains a high hyperforin content and Rebalance® because it is controlled for a low hyperforin content. In silico analysis revealed a weak hyperforin-rPXR binding affinity, which was further supported in cell-based reporter gene assays showing no hyperforin-mediated reporter activation in presence of rPXR. However, cellular exposure to Hyperiplant® and Rebalance® transactivated the CYP3A-reporter 3.8 fold and 2.8 fold, respectively, and they induced Cyp3a23-3a1 mRNA expression in rat hepatoma cells compared to control 48-fold and 18-fold, respectively. In Wistar rats treated for 10 days with 400 mg/kg of Hyperiplant® we observed 1.8-times the Cyp3a23-3a1 mRNA expression, a 2.6-fold higher CYP3A23-3A1 protein amount and a 1.6-fold increase in activity compared to control. For Rebalance® we only observed a 1.8-fold hepatic increase of CYP3A23-3A1 protein compared to control animals. Even though there are differing effects on rCyp3a23-3a1/CYP3A23-3A1 in rat liver reflecting the hyperforin content of the SJW extracts, the modulation is most likely not linked to an interaction of hyperforin with rPXR. Significance Statement Treatment with St John's wort (SJW) has been reported to affect CYP3A expression and activity in rats. Our comparative study further supports this finding but shows that the PXR-ligand hyperforin is not the driving force for changes in rat CYP3A23-3A1 expression and function in vivo and in vitro Importantly, CYP3A induction mimics findings in humans but our results suggest that another so far unknown constituent of SJW is responsible for the expression and function modifying effects in rat liver.

How Changes in Nitrogen Levels Impact Metabolites and Physiological Features of Hypericum androsaemum

Background: The manipulation of mineral nutrition may affect secondary metabolite production and thus determine the nutritional value of food/medicinal plants. Objectives: The present study investigated the impact of nitrogen supply on the production of secondary metabolites and physiological parameters of Hypericum androsaemum plants grown in sand culture. Methods: Four-week plants were irrigated by full-strength Hoagland solution (control), nitrate was reduced to 1/3, and no nitrate was provided for 14 days. Results: The growth of shoots was inhibited by nitrate limitation, but root growth was stimulated. Soluble proteins and water content were significantly affected by nitrate deficiency. Antioxidative enzyme activities (ascorbate peroxidase (APX) and superoxide dismutase (SOD)) were increased due to nitrate deficiency. Treatment of plants with NO3- deficiency caused an increase in total soluble phenols and phenylalanine ammonia-lyase (PAL) activity, while flavonoid content was mainly elevated by NO3- deficiency. At the level of specific metabolites, NO3- deficiency positively affected hypericin and pseudohypericin content but decreased hyperforin content. The expression of the putative gene for hypericin biosynthesis (hyp-1) was up-regulated in response to nitrate deficiency. Conclusions: The results demonstrated that the nitrogen deficiency in the H. androsaemum plant positively affected the antioxidant system, valuable phenols, and secondary metabolites.

Cytotoxic and Antioxidant Activity of Hypericum perforatum L. Extracts against Human Melanoma Cells from Different Stages of Cancer Progression, Cultured under Normoxia and Hypoxia.

Oxidative stress and the hypoxic microenvironment play a key role in the progression of human melanoma, one of the most aggressive skin cancers. The aim of our study was to evaluate the effect of Hypericum perforatum extracts of different origins (both commercially available (HpEx2) and laboratory-prepared from wild grown (HpEx12) and in vitro cultured (HpEx13) plants) and hyperforin salt on WM115 primary and WM266-4 lymph node metastatic human melanoma cells cultured under normoxic and hypoxic conditions. The polyphenol content, radical scavenging activity, and hyperforin concentration were determined in the extracts, while cell viability, apoptosis, ROS production, and expression of NRF2 and HO-1, important oxidative stress-related factors, were analyzed after 24 h of cell stimulation with HpExs and hyperforin salt. We found that cytotoxic, pro-apoptotic and antioxidant effects depend on the extract composition, the stage of melanoma progression, and the oxygen level. Hyperforin salt showed lower activity than H. perforatum extracts. Our study for the first time showed that the anticancer activity of H. perforatum extracts differs in normoxia and hypoxia. Importantly, the composition of extracts of various origins, including in vitro cultured, resulting in their unique properties, may be important in the selection of plants for therapeutic application.

Antioxidant potential and essential oil properties of Hypericumperforatum L. assessed by application of selenite and nano-selenium

It is necessary to develop a simple way to achieve food quality quantitatively. Nanotechnology is a key advanced technology enabling contribution, development, and sustainable impact on food, medicine, and agriculture. In terms of medicinal and therapeutic properties, Hypericumperforatum is an important species. For this study, a randomized complete block design with three replications was used in each experimental unit. The foliar application of selenite and nano-selenium (6, 8, 10, and 12 mg/l), control (distilled water), at the rosette stage and harvesting at 50% flowering stage has been applied as an alleviation strategy subjected to producing essential oils and antioxidant activity. Experimental results revealed that the selenite and nano selenium fertilizers had a significant effect on traits such as total weight of biomass, essential oil percentage, the content of hypericin and hyperforin, the selenium accumulation in the plant, relative leaf water content, chlorophylls, phenolic content, proline, catalase, peroxidase, malondialdehyde, and DPPH. The highest essential oil content was obtained from the control treatment when the accumulation of selenium was achieved with 12 mg/l nano-selenium. The maximum rate of hypericin was seen in the foliar application of 8 mg/l selenite whereas the maximum hyperforin was gained at 10 mg/l selenium. Conceding that the goal is to produce high hypericin/ hyperforin, and also the accumulation of selenium in the plant, treatments of 6 and 8 mg/l of selenite and nano-selenium could be applied. Consequently, an easy detection technique proposed herein can be successfully used in different ranges, including biology, medicine, and the food industry.

Effects of light intensity and LED spectrum on the medicinal component accumulation of hydroponic Hypericum perforatum L. under controlled environments

Medicinal components of Hypericum perforatum L. plants varies widely due to fluctuations in growth environment and biotic and abiotic contamination during cultivation management. The quality of extracts or preparations is difficult to control because of the unstable raw materials. The aim of this study is to enhance the yield and medicinal component contents of H. perforatum by optimizing lighting factors under controlled environment. H. perforatum plants were hydroponically cultivated for 30 d under 3 levels of photosynthetic photon flux density (PPFD) with 200, 300, and 400 μmol/(m2·s) using white LEDs (R:B ratio is the ratio of red light to blue light, R:B ratio of 0.9 and 1.8) and white plus red LED (R:B ratio of 2.7). The results showed that PPFD and LED spectrum had significant effects on the growth and accumulation of medicinal components of H. perforatum. Biomass accumulation of stem, leaf, and root increased linearly with the increase of PPFD under each LED spectrum. Fresh weights and dry weights of stem, leaf, and root were significantly higher under a PPFD of 400 μmol/(m2·s) with R:B ratio of 0.9 than those of 200 μmol/(m2·s), respectively. The relative growth rate and net photosynthetic rate showed linear relationships with PPFD under the same LED spectrum. Total hypericin content, total hyperforin content, and energy yield of hypericin increased with increasing PPFD. Total hypericin content and energy yield of hypericin of P400-L0.9 were 78% and 89% more than those of P400-L2.7, respectively. Total hyperforin content and energy yield of hyperforin of P400-L0.9 and P400-L2.7 were no significant differences. Based on energy efficiency, an R:B ratio of 0.9 of white LEDs with a PPFD of 400 μmol/(m2·s) was beneficial to improve medicinal component contents of hydroponic H. perforatum in plant factory with LED lighting. Keywords: LED spectrum, light intensity, controlled environments, Hypericum perforatum L. DOI: 10.25165/j.ijabe.20221505.7373 Citation: Gao W, He D X, Zheng J F, Li Y. Effects of light intensity and LED spectrum on the medicinal component accumulation of hydroponic Hypericum perforatum L. under controlled environments. Int J Agric & Biol Eng, 2022; 15(5): 63–69.

Red LED light promotes biomass, flowering and secondary metabolites accumulation in hydroponically grown Hypericum perforatum L. (cv. Topas)

To increase the crop yield and quality, the plant cultivation in equipped growing systems is an alternative to open field production. Artificial light sources are a critical component in the plant factory systems and red and blue wavelengths have a powerful impact on plant growth and photosynthetic capacity. In this study, the effect of different photon flux ratios of red (R) light to blue (B) LED light including 100 R:0B, 80 R:20B, 50 R:50B, 20 R:80B, 0 R:100B and white LEDs (as control) were evaluated on the growth, flowering and secondary metabolite contents of Hypericum perforatum L. cv. Topas at 150 µmol m−2 s−1 irradiation for a 16/8 h.d‐1 photoperiod. Results showed that the plant growth characteristics such as fresh and dry weight, leaf area and number, plant height, flower number and diameter were significantly (P < 0.05) maximum under the enhancement of R/B ratio, especially under 100% red light treatment. Despite the higher transpiration and net photosynthesis rate, photosynthetic pigment concentration, stomatal surface area and RuBisCO activity, the plants under the high fraction of blue light showed the poorest growth and flowering traits. The maximum content of hypericin (14.24 mg g‐1 DW), pseudohypericin (25.98 mg g−1 DW) and hyperforin (163.06 mg g−1 DW) was obtained in flower under 100% red LED. Antioxidant enzymes activities (CAT and POD), total phenolic and flavonoid content were also higher in plants exposed to the mixture of red and blue light. Plants grown under white LED were mediocre in terms of growth parameters and secondary metabolites. These findings showed that the 100% red LED light is suitable for H. perforatum plant growth, flowering, as well as secondary metabolites accumulation in the plant factory system.

Development of new and validated HPTLC methods for the qualitative and quantitative analysis of hyperforin, hypericin and hyperoside contents in Hypericum species

Hypericum perforatum L. (St. John's wort) is a medicinally important member of Hypericaceae. Many pharmacological activities have been mostly attributed to its hyperforin, hypericin and/or hyperoside contents. Therefore, qualitative and quantitative determinations of these ingredients are essential to justify the beneficial effects of St. John's wort on health. In the European Pharmacopoeia, the TLC and HPLC methods were given for this purpose. High performance thin layer chromatography (HPTLC) has recently become increasingly used as a suitable technique for analysing herbal drugs. This study aims to develop new and validated HPTLC methods to analyse these active components in different Hypericum spp. to find other suitable species to replace the official plant. Three different mobile phases were developed: n-hexane-ethyl acetate (8:2) for hyperforin analysis, toluene-chloroform-ethyl acetate-formic acid (8:5:3.5:0.6) for hypericin analysis and ethyl acetate-formic acid-acetic acid-water (15:2:2:1) for hyperoside analysis. These newly developed and validated HPTLC systems were further applied to determine their concentrations in different Hypericum species. Hyperforin concentration was found between 6.40 to 26.40 mg/g only in H. triquetrifolium, H. scabrum and two H. perforatum samples; hypericin was detected between 0.81 and 1.41 mg/g only in H. bithynicum, H. perfoliatum, H. triquetrifolium and two H. perforatum samples; and hyperoside was identified in all tested specimens ranging from 1.01 to 9.73 mg/g. The new HPTLC methods developed and validated in the present study may ensure reliable results for the qualification and quantification of hyperforin, hypericin and hyperoside contents in Hypericum species.

A comparison of artificial intelligence techniques for predicting hyperforin content in Hypericum perforatum L. in different ecological habitats.

Hyperforin, a major bioactive constituent of Hypericum concentration, is impacted by various phenological phases and soil characteristics. We aimed to design a model predicting hyperforin content in Hypericum perforatum based on different ecological and phenological conditions. We employed artificial intelligence modeling techniques including multilayer perceptron (MLP), radial basis function (RBF), and support vector machine (SVM) to examine the factors critical in predicting hyperforin content. We found that the MLP model (R 2 = .9) is the most suitable and precise model compared with RBF (R 2 = .81) and SVM (R 2 = .74) in predicting hyperforin in H. perforatum based on ecological conditions, plant growth, and soil features. Moreover, phenological stages, organic carbon, altitude, and total N are detected in sensitivity analysis as the main factors that have a considerable impact on hyperforin content. We also report that the developed graphical user interface would be adaptable for key stakeholders including producers, manufacturers, analytical laboratory managers, and pharmacognosists.

Prescription of St John's Wort extracts: clinical risks due to differences in their hyperforin content

Prescription of St John's Wort extracts: clinical risks due to differences in their hyperforin content

Harvest time affects antioxidant capacity, total polyphenol and flavonoid content of Polish St John\u2019s wort\u2019s (Hypericum perforatum L.) flowers

The polyphenol content and antioxidant capacity of hyperforin and hypericin-standardized H. perforatum L. extracts may vary due to the harvest time. In this work, ethanol and ethanol–water extracts of air-dried and lyophilized flowers of H. perforatum L., collected throughout a vegetation season in central Poland, were studied. Air-dried flowers extracts had higher polyphenol (371 mg GAE/g) and flavonoid (160 mg CAE/g) content, DPPH radical scavenging (1672 mg DPPH/g), ORAC (5214 µmol TE/g) and FRAP (2.54 mmol Fe2+/g) than lyophilized flowers extracts (238 mg GAE/g, 107 mg CAE/g, 1287 mg DPPH/g, 3313 µmol TE/g and 0.31 mmol Fe2+/g, respectively). Principal component analysis showed that the collection date influenced the flavonoid and polyphenol contents and FRAP of ethanol extracts, and DPPH and ORAC values of ethanol–water extracts. The ethanol extracts with the highest polyphenol and flavonoid content protected human erythrocytes against bisphenol A-induced damage. Both high field and benchtop NMR spectra of selected extracts, revealed differences in composition caused by extraction solvent and raw material collection date. Moreover, we have shown that benchtop NMR can be used to detect the compositional variation of extracts if the assignment of signals is done previously.

Protective Role of St. John's Wort and Its Components Hyperforin and Hypericin against Diabetes through Inhibition of Inflammatory Signaling: Evidence from In Vitro and In Vivo Studies.

Diabetes mellitus is a very common chronic disease with progressively increasing prevalence. Besides the well-known autoimmune and inflammatory pathogenesis of type 1 diabetes, in many people, metabolic changes and inappropriate lifestyle favor a subtle chronic inflammatory state that contributes to development of insulin resistance and progressive loss of β-cell function and mass, eventually resulting in metabolic syndrome or overt type 2 diabetes. In this paper, we review the anti-inflammatory effects of the extract of Hypericum perforatum L. (St. John’s wort, SJW) and its main active ingredients firstly in representative pathological situations on inflammatory basis and then in pancreatic β cells and in obese or diabetic animal models. The simultaneous and long-lasting inhibition of signal transducer and activator of transcription (STAT)-1, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and mitogen-activated protein kinases (MAPKs)/c-jun N-terminal kinase (JNK) signaling pathways involved in pro-inflammatory cytokine-induced β-cell dysfunction/death and insulin resistance make SJW particularly suitable for both preventive and therapeutic use in metabolic diseases. Hindrance of inflammatory cytokine signaling is likely dependent on the hyperforin content of SJW extract, but recent data reveal that hypericin can also exert relevant protective effects, mediated by activation of the cyclic adenosine monophosphate (cAMP)/protein kinase cAMP-dependent (PKA)/adenosine monophosphate activated protein kinase (AMPK) pathway, against high-fat-diet-induced metabolic abnormalities. Actually, the mechanisms of action of the two main components of SJW appear complementary, strengthening the efficacy of the plant extract. Careful quantitative analysis of SJW components and suitable dosage, with monitoring of possible drug–drug interaction in a context of remarkable tolerability, are easily achievable pre-requisites for forthcoming clinical applications.

Optimization of Light Intensity, Temperature, and Nutrients to Enhance the Bioactive Content of Hyperforin and Rutin in St. John's Wort.

St. John’s wort (Hypericum perforatum L.) is a medicinal plant that alleviates depression and other disorders due to its abundance of active ingredients. Hyperforin, rutin, and melatonin are the main active, and important, ingredients in St. John’s wort that alleviate depression. In order to investigate the optimal conditions for accumulating these active ingredients, design of experiments and response surface methodology (RSM) was employed in this study. Two-month-old St John’s wort plants were cultivated in growth chambers at varying temperatures, light intensities, and nutrient solution concentrations before analysis by HPLC, for determining differences in hyperforin, rutin, and melatonin content. The results showed that hyperforin and rutin contents were significantly influenced by temperature (18–23 °C) and light intensity (49–147 μmol m−2 s−1 photosynthetic photon flux density (PPFD)), whereas Hoagland’s nutrient solution concentration (25–75%) had little effect. The accumulation of melatonin might not be influenced by cultivation conditions. Light intensity and temperature are easily controlled environmental factors in artificial cultivation, both of which are related to secondary metabolite production in the plant. Based on RSM, the optimal conditions for the accumulation of hyperforin and rutin were obtained. The maximum content of hyperforin was 5.6 mg/g, obtained at a temperature of 19 °C, a nutrient solution concentration of 45%, and a light intensity of 49 μmol m−2 s−1 PPFD. The maximum content of rutin was 3.8 mg/g obtained at a temperature of 18 °C, a nutrient solution concentration of 50%, and a light intensity of 147 μmol m−2 s−1 PPFD. This evaluation of suitable conditions for the accumulation of bioactive compounds in St. John’s wort can be applied to plant factories on a large scale.

STABILITY OF SOME BIOLOGICALLY ACTIVE SUBSTANCES IN EXTRACTS AND PREPARATIONS BASED ON ST. JOHN’S WORT (HYPERICUM PERFORATUM L.) AND SAGE (SALVIA OFFICINALIS L.)

Quality, authenticity and stability control of medicinal plant materials and preparations based on them is an important challenge in industries using substances and materials derived from plants. St. John’s wort (Hypericum perforatum L.) and sage (Salvia officinalis L.) are one of the most common and best-selling medicines in the world. They are composed of phloroglucinols and diterpenes possessing pharmacological properties. However, some of the compounds (hyperforin, carnosic acid, etc.) determining these qualities decompose under the influence of various physical and chemical factors with the formation of stable and unstable intermediates. Considering the importance of utilization of these plants, the stability of hyperforin, carnosic acid and other compounds in their extracts and preparations during extraction and storage was studied in this work. Extracts and preparations were studied by high performance liquid chromatography (HPLC) with diode-array and mass spectrometric detection. Plant extracts were obtained by heating (HE) as well as using ultrasonic (USE), microwave (MWE) and supercritical (SSE) extraction. The stability of hyperforins, their degradation, as well as the relationship between hyperforin and furohyperforin contents in extracts and preparations based on H. perforatum L. during storage were studied. Hyperforin to furohyperforin ratio in the extracts ranged from 18 to 5 depending on the origin of the medicinal plant material. The stability and transformation of carnosic acid was evaluated, the relationship between carnosic acid and carnosol contents and conditions of sample preparation and storage as well as industrially manufactured preparations based on S. officinalis L. was revealed. The possibility of applicability of hyperforin to furohyperforin ratio in H. perforatum L. and carnosic acid to carnosol ratio in S. officinalis L. was shown to confirm the quality of extracts and preparations on their basis. Since rutin to hyperforin ratio in H. perforatum L. is constant, this indicator can be used to confirm the quality and authenticity of the feedstock, regardless of where the plant grows.

Clinical relevance of St. John's wort drug interactions revisited

The first clinically relevant reports of preparations of St. John's wort (SJW), a herbal medicine with anti‐depressant effects, interacting with other drugs, altering their bioavailability and efficacy, were published about 20 years ago. In 2000, a pharmacokinetic interaction between SJW and cyclosporine caused acute rejection in two heart transplant patients. Since then, subsequent research has shown that SJW altered the pharmacokinetics of drugs such as digoxin, tacrolimus, indinavir, warfarin, alprazolam, simvastatin, or oral contraceptives. These interactions were caused by pregnane‐X‐receptor (PXR) activation. Preparations of SJW are potent activators of PXR and hence inducers of cytochrome P450 enzymes (most importantly CYP3A4) and P‐glycoprotein. The degree of CYP3A4 induction correlates significantly with the hyperforin content in the preparation. Twenty years after the first occurrence of clinically relevant pharmacokinetic drug interactions with SJW, this review revisits the current knowledge of the mechanisms of action and on how pharmacokinetic drug interactions with SJW could be avoided.Linked ArticlesThis article is part of a themed section on The Pharmacology of Nutraceuticals. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v177.6/issuetoc

Toward enhanced hyperforin production in St. John's wort root cultures.

During the past decades, several trials targeted a stable, sustainable and economic production of St. John's wort (Hypericum perforatum) extract. The value of this extract stems from its use to treat depression and skin irritation due to its hyperforin content. Previously, hyperforin-forming in vitro root cultures were established. Here, detailed growth and production kinetics have been analyzed over 40 days of cultivation. In the first 10 days, sucrose was completely hydrolyzed to glucose and fructose. The ammonium consumption supported the increase in the biomass and hyperforin production. When sucrose was replaced with glucose/fructose, the linear growth phase started 6 days earlier and resulted in a higher space-time-yield. The maximum hyperforin production was 0.82mgL-1 day-1, which was 67 % higher than in the sucrose-supplemented standard cultivation. Buffering the sucrose-supplemented medium with phosphate caused a 2.7-fold increase in the product to biomass yield coefficient. However, the combination of monosaccharides and buffering conditions did not cause an appreciable improvements in the production performance of the shake flask approaches. A potential scalability from flask to lab-scale stirred bioreactors has been demonstrated. The results obtained offer a basis for a scalable production of hyperforin and a sustainable source for a tissue culture-based phytomedicine.

Physiologically Based Pharmacokinetic Modelling of Hyperforin to Predict Drug Interactions with St John's Wort.

Herb-drug interactions with St John's wort (SJW) have been widely studied in numerous clinical studies. The objective of this study was to develop and evaluate a physiologically based pharmacokinetic (PBPK) model for hyperforin (the constituent of SJW responsible for interactions), which has the potential to provide unique insights into SJW interactions and allow prediction of the likely extent of interactions with SJW compared to published interaction reports. A PBPK model of hyperforin accounting for the induction of cytochrome P450 (CYP) 3A, CYP2C9 and CYP2C19 was developed in the Simcyp® Simulator (version 17) and verified using published, clinically observed pharmacokinetic data. The predictive performance of this model based on the prediction fold-difference (expressed as the ratio of predicted and clinically observed change in systemic exposure of drug) was evaluated across a range of CYP substrates. The verified PBPK model predicted the change in victim drug exposure due to the induction by SJW (expressed as area under the plasma concentration-time curve (AUC) ratio) within 1.25-fold (0.80-1.25) of that reported in clinical studies. The PBPK simulation indicated that the unbound concentration of hyperforin in the liver was far lower than in the gut (enterocytes). Simulations revealed that induction of intestinal CYP enzymes by hyperforin was found to be more pronounced than the corresponding increase in liver CYP activity (15.5- vs. 1.1-fold, respectively, at a hyperforin dose of 45mg/day). In the current study, a PBPK model for hyperforin was successfully developed, with a predictive capability for the interactions of SJW with different CYP3A, CYP2C9 and CYP2C19 substrates. This PBPK model is valuable to predict the extent of herb-drug interactions with SJW and help design the clinical interaction studies, particularly for new drugs and previously unstudied clinical scenarios.

Hyperforin Induces Apoptosis Through Extrinsic/Intrinsic Pathways and Inhibits NF-ĸB-modulated Survival and Invasion Potential in Bladder Cancer.

Muscle-invasive bladder cancer (MIBC) has long been recognized as a difficult to treat cancer type, thus a new treatment strategy is needed. The major purpose of the present study was to verify the anticancer effect of hyperforin and the mechanism through which it affects tumor cell growth and invasion in bladder cancer in vitro. Bladder cancer TSGH-8301 cells were treated with different concentrations of hyperforin for different durations of time. The changes in cell viability, production of calcium and reactive oxygen species (ROS), and anti-apoptotic signaling were evaluated using MTT assay, flow cytometry, and western blot analysis. The effect of hyperforin on the expression of nuclear factor-kappaB (NF-ĸB) p65 (Ser276), tumor progression-associated proteins, as well as on cell invasion was investigated using western blotting and cell invasion assay, respectively. Hyperforin significantly induces apoptosis, extrinsic/intrinsic apoptotic signaling, accumulation of cytosol ROS, and calcium signalling. Hyperforin also significantly diminishes the expression of NF-ĸB p65 (Ser276), anti-apoptotic and tumor progression-associated proteins, as well as the cell invasion ability of TSGH-8301 cells. Our findings demonstrate that hyperforin triggers apoptosis depending on extrinsic/intrinsic pathways and suppresses NF-ĸB-mediated cell survival as well as the invasive properties of bladder cancer in vitro.

Hyperforin-Induced Activation of the Pregnane X Receptor Is Influenced by the Organic Anion-Transporting Polypeptide 2B1.

The herbal remedy St. John's wort (SJW) is used in the treatment of mild depressive symptoms and is known for its drug-drug interaction potential when enhanced expression of CYP3A4 modifies clearance of concomitantly applied substrate drugs. Hyperforin is one constituent of SJW that alters CYP3A4 expression by activation of the nuclear receptor pregnane X receptor (PXR). However, little is known about the transmembrane transport of hyperforin. One membrane protein that modulates cellular entry of drugs is the organic anion-transporting polypeptide (OATP) 2B1. It was the aim of this study to test whether hyperforin interacts with this transport protein. Transport inhibition studies and competitive counterflow experiments suggested that hyperforin is a substrate of OATP2B1. This notion was validated by showing that the presence of OATP2B1 enhanced the hyperforin-induced PXR activation in cell-based luciferase assays. Moreover, in Caco-2 cells transcellular transport of the known OATP2B1 substrate atorvastatin was changed in the presence of hyperforin, resulting in an increased efflux ratio. Eleven commercially available SJW formulations were assessed for their influence on OATP2B1-mediated transport of estrone 3-sulfate and for their impact on CYP3A4 promoter transactivation. The correlation between effect size and the hyperforin content as determined by high-performance liquid chromatography with ultraviolet detection suggested that hyperforin is the major determinant. Our results indicate an interaction between hyperforin and OATP2B1, which is not only known to contribute to hepatocellular uptake but also to intestinal absorption of its substrates. These findings extend the complexity of mechanisms that should be considered when evaluating the interaction potential of SJW preparations.