09/07/2021

1. 1. Double-blind placebo controlled trial of the anxiolytic effects of a standardized Echinacea extract
      Abstract: Earlier studies suggested that specific Echinacea preparations might decrease anxiety. To further  study the issue, we performed a double blind, placebo controlled trial with a standardized Echinacea angustifolia root extract. Participants were volunteers scoring above 45 points on the state or on the trait subscale of the State Trait Anxiety Inventory (STAI). They were treated with 40 mg Echinacea or with placebo tablets twice daily for 7 days followed by a 3 week‐long washout period. Participants were also administered the Beck Depression Inventory (BDI) and the Perceived Stress Scale (PSS). In the Echinacea group, state anxiety scores decreased by approximately 11 points by the end of the treatment period, whereas the decrease was around 3‐points in the placebo group (p< 0.01). The effect maintained over the washout period. The difference from placebo was significant from the 7th day of treatment throughout. Changes were less robust with trait anxiety scores, but the preparation performed better than placebo in patients with high baseline anxiety. Neither BDI nor PSS scores were affected by the treatments. Adverse effects were rare and mild, and all were observed in the placebo group. These findings suggest that particular Echinacea preparations have significant beneficial effects on anxiety in humans. [Haller J, Krecsak L, Zambori J. Double-blind placebo controlled trial of the anxiolytic effects of a standardized Echinacea extract. Phytotherapy Research 2019;1-9.]
   2. The Anxiolytic Potential and Psychotropic Side Effects of an Echinacea Preparation in Laboratory Animals and Healthy Volunteers. We investigate that toxicity, psychotropic side effects and anxiolytic potential of and Echinacea angustifolia extract that produced promising effects in laboratory tests performed earlier. Rats were studied in the elevated plus-maze, conditioned fear, open-field, object recognition and conditioned place preference test. Toxicity was studied in rats after intragastric administration. The preparation decreased anxiety in the elevated plus-maze and ameliorated contextual conditioned fear. No lethality or behavioral signs of discomfort were noticed in rats treated with 1000 and 3000mg/kg Echinacea angustifolia. The extract was without effect in tests of locomotion (open-field), memory (object recognition) and rewarding potential (conditioned place preference) within a wide dose range. A pharmacological formulation based on the same Echinacea angustifolia extract was tested in human subjects. One or two tablets per day were administered for 1 week to healthy volunteers scoring high on the State-Trait Anxiety Inventory (STAI). The tablets contain 20mg of the plant extract. Data were collected using a structured self-assessment diary technique. The high dose (2 tablets per day) decreased STAI scores within 3 days in human subjects, an effect that remained stable for the duration of the treatment (7 days) and for the 2 weeks that followed treatment. The lower dose (1 tablet per day) did not affect anxiety significantly. [Haller J, et al. The Anxiolytic Potential and Psychotropic Side Effects of an Echinacea Preparation in Laboratory Animals and Healthy Volunteers. Phytother Res. 2013 Jan;27(1):54-61.]
   3. An investigation into the anxiety-relieving and mood-enhancing effects of Echinacea
      angustifolia (EP107™): A randomised, double-blind, placebo-controlled study. Background: The acute anxiolytic effects of the echinacea angustifolia extract (EP107TM) have been demonstrated in two previous human trials. The goals of this study were to
      examine the anxiolytic and mood-enhancing effects of echinacea angustifolia over a longer duration with a larger sample size. Methods: In this 6-week, 3-arm, parallel-group, doubleblind, randomised controlled trial, 108 adults with mild-to-moderately severe anxiety were recruited and randomised to receive either a placebo, 20 mg, or 40 mg of echinacea angustifolia, twice daily. Outcome measures included the Clinically Useful Anxiety Outcome Scale (CUXOS), Positive and Negative Affect Scale (PANAS), Short Form-36 (SF-36), and Bergen Insomnia Scale (BIS). Results: Based on data collected from 104 participants, both doses of echinacea were associated with overall reductions in anxiety, although improvements were not different from the placebo. However, both doses of echinacea were associated with greater improvements in the PANAS positive and negative affect scores, and SF-36 emotional wellbeing score compared to the placebo. Limitations: The positive improvements associated with echinacea were only identified via the secondary outcome measures and, therefore, require validation in future trials. Conclusions: The echinacea angustifolia extract (EP107TM) administered for 6 weeks at a dose of 40 and 80 mg daily was not associated with greater improvements in anxiety in adults with mild-to-moderately severe anxiety compared to the placebo. However, there were greater improvements in positive and
      negative affect, and emotional wellbeing, suggesting antidepressant effects. Further studies using clearly-defined populations and validated outcome measures will be useful in future trials. [Lopresti AL, Smith SJ. An investigation into the anxiety-relieving and moodenhancing effects of Echinacea angustifolia (EP107™): A randomised, double-blind, placebo-controlled study. J Affect Disord. 2021;293:229-237.]
   4. Possible Role of Fat Tissue in the pharmacokinetics of Dodeca-2E, 4E, 8Z, 10E/Ztetraenoic Acid Isobutylamides after Oral Administration of Echniacea angustifolia Extract in Rats. Alkamides are one of the most important constituents of lipophilic extracts of Echinacea angustifolia roots. These compounds play an important role in the versatile pharmacological actions of this plant. The present study aimed to compare the concentrations of isomeric dodeca-2E,4E,8Z,10E/Z-tetraenoic acid isobutylamides (DTAI) in brain and periepididymal fat tissues and blood plasma of rats. Thirty, 60, 240 and 720 min after the oral administration of E. angustifolia root extract, tissue and plasma concentrations were determined by reversed-phase HPLC with ESI-MS/MS detection. The calculated terminal t of the mixture of DTAI was 8.28 h, which indicates a relatively slow elimination. In the 0.5–4 h period the brain/plasma and fat/plasma concentration ratios were continuously above 3 and 18, respectively, followed by equilibrium at 12 h. Our results indicate substantial accumulation of alkamides in lipid-rich tissues, which presumably contributes to a maintained pharmacological action. [Jedlinszki N, Redei D, Haller J, Freund T, Hohmann J, Zupko I. Possible Role of Fat Tissue in the Pharmacokinetics of Dodeca-2E, 4E, 8Z, 10E/Z-tetraenoic Acid Isobutylamides after Oral Administration of Echinacea angustifolia Extract in Rats. Natural Products Communications, Volume 9 Issue 6:737-888]
   5. The Effect of Echinacea Preparations in Three Laboratory Tests of Anxiety: Comparison with Chlordiazepoxide. Echinacea preparations are traditionally used to treat upper respiratory infections and inflammation. No psychotropic effects of Echinacea have been reported so far, although some recently reported active constituents are behaviorally active. Prompted by these finding, the anxiolytic potential of five different Echinacea preparations was evaluated. Three of these decreased anxiety but two of them had a very narrow effect dose range. Only one extract decreased anxiety within a wide dose-range (3-8mg/kg). Anxiolytic effects were consistently seen in three different tests of anxiety, the elevated plus-maze, social interaction and shock-induced social avoidance test. No locomotor suppressant effects were seen at nay dose. Noteworthy, the doses that showed anxiolytic effects in the present study were much lower than those used in the laboratory models of the traditional indications; Chlordiazepoxide robustly decreased anxiety-like behavior in all texts but suppressed locomotion at higher doses. Perceived and real risks of conventional medications increase the demand for alternative therapies, provided that these are safe and efficient. Earlier evidence shows that Echinacea preparations have an excellent safety profile, where our finding suggest for the first time that certain preparations have a considerable anxiolytic potential. Further research is required to identify facts that differentiate efficient and inefficient preparations. [Haller J, et al. The Effect of Echinacea Preparations in Three Laboratory Tests of Anxiety: Comparison with Chlordiazepoxide. Phytother Res. 2010 Nov;24(11):1605-13.]
   6. The Effects of an Echinacea Preparation on Synaptic Transmission and the Firing Properties of CA1 Pyramidal Cells in the Hippocampus. Traditionally, Echinacea preparations are used as anti-inflammatory agents and immune-enhancers. In addition to these effects, their anxiolytic potency has been recognized recently in laboratory tests. Our aim in this study was to uncover the potential effects of an Echinacea preparation on neuronal operations in the hippocampus, a brain region this is involved in anxiety and anxiety-related behaviors. Using in vitro electrophysiological techniques, we observed that excitatory synaptic transmission is hippocampal slices was significantly suppressed by an Echinacea extract found to be effective in anxiety tests. In contrast, no change in inhibitory synaptic transmission could be detected upon application on this extract. In addition, our experiments revealed that at low concentration the Echinacea extract reduced the spiking activity of CA1 pyramidal cells, while at high concentration increased it. This later observation was parallel to the reduction in the magnitude of the h-current-mediated voltage responses in pyramidal cells. At any concentrations, the passive membrane properties of CA1 pyramidal cells were found to be unaltered by the Echinacea extract. In summary, the Echinacea extract can significantly regulate excitatory, but not inhibitory, synaptic transmission in the hippocampus, and this action might be involved in its anxiolytic effects observed in behavior tests. [H jos N, et al. The Effects of an Echinacea Preparation on Synaptic Transmission and the Firing Properties of CA1 Pyramidal Cells in the Hippocampus. Phytother Res. 2012 Mar;26(3):354-62.]
   7. Alkamides and a neolignan from Echinacea purpurea roots and the interaction of alkamides with G-protein-coupled cannabinoid receptors. Multiple chromatographic separations of the CHCL3-soluble extract of the roots of Echinacea purpurea led to the isolation of 19 compounds. Four natural products, three alkamides and nitidanin diisovalerianate, were identified, and five further compounds were detected for the first time in the spices. Additionally, 10 known Echinacea purpurea metabolites were isolated. The structures were determined by mass spectrometry and advanced 1D and 2D NMR techniques. The bioactivity of the isolated compounds was studied in the [35S] GTPγS-binding experiments performed on rat brain membrane preparations. Both partial and inverse agonist compounds for cannabinoid (CB1) receptors were identified among the metabolites, characterized by weak to moderate interactions with the G-protein signaling mechanisms. The G-protein-modulating activities of the Echinacea compounds are rather far from the full agonist effects seen with the CB1 receptor agonist reference compound arachidonyl-2’-chloroethylamide (ACEA). However, upon coadministration with ACEA, a number of them proved capable of inhibiting the stimulation of the pure agonist, thereby demonstrating cannabinoid receptor antagonist properties. [Hohmann J, et al. Alkamides and a neolignan from Echinacea purpurea roots and the interaction of alkamides with G-protein-coupled cannabinoid receptors. Phytochemistry 2011 Oct;72(14-15):1848-53.]