Anxiolytic activity of ferulic acid in the light-dark test in zebrafish

The anxiety disorders belong to a group of mental disorders in which the patients present excessive fear and worry. Studies with ferulic acid have shown positive results on treating depressive symptoms. As many antidepressive drugs are effective in treating anxiety, the objective of the present study was to evaluate ferulic acid’s anxiolytic activity and possible mechanism of action in the light-dark test in zebrafish. To evaluate anxiolytic activity, the light-dark preference test was performed after exposure of the animals to ferulic acid or positive control (clonazepam or fluoxetine). Ferulic acid increased the time spent in the clear compartment at concentrations of 250 and 500 mg/L, not differing from the groups exposed to clonazepam or fluoxetine. To evaluate the possible mechanism of action, pre-exposure to flumazenil was carried out, followed by exposure to ferulic acid or positive control, with subsequent testing. Pre-exposure to flumazenil caused a significant reduction in the time spent in the clear compartment of ferulic acid and clonazepam groups but did not alter the effect of exposure to fluoxetine. These results suggest that ferulic acid promotes an anxiolytic effect, possibly through an action at the benzodiazepine binding site at the GABAA receptor.

STUDY OF THE ANXIOLYTIC ACTIVITY OF ETHANOLIC EXTRACT OF ROOT OF ACORUS CALAMUSIN ALBINO MICE USING MIRRORED CHAMBER

Objective: Anxiety is classified as a form of sub-acute or chronic fear. Root of Acorus calamus has been traditionally used as an anxiolytic. The aim of the study is to assess the anxiolytic activity of ethanolic extract of the root of Acorus calamus (EEAC) by Mirrored Chamber Test on Albino mice. Methods: Albino mice of either sex were taken and divided into five groups, each consisting of 5 mice. One group was used as control, one as standard (Diazepam) and three as test groups treated with 100, 200 & 400 mg/kg of EEAC. The drugs i.e., 10 ml/kg of Normal saline for control, Diazepam 2 mg/kg (standard) and 100, 200 & 400 mg/kg EEAC (test groups) were injected intraperitoneally (i.p.), 30 mins before placing them in the center of the mirrored chamber. Latency to enter the chamber, number of entries in five minutes and total time spent in the chamber during 5 minutes test period were compared in each group. Data were statistically analyzed by one-way analysis of variance followed by multiple Dunnett’s test. Results: EEAC showed significant increase in all the parameters for the doses of 100, 200 and 400 mg/kg in a dose dependent manner. Conclusion: EEAC has anxiolytic activity.

Design, synthesis, and pharmacological activity of new pyrrolo[1,2-a]pyrazine translocator protein (tspo) ligands

Background: Translocator protein 18 kDa (TSPO) is a promising target for the creation of effective and safe neuropsychotropic drugs. The ligands of TSPO exhibit anxiolytic, antidepressant, neuroprotective and other activities without the side effects of benzodiazepines. Methods: New TSPO ligands in the series of N,1-diphenylpyrrolo[1,2-a]pyrazine-3-carboxamides derivatives were designed using calculated pharmacophore model and molecular docking analysis. The synthesis of new compounds was carried out by two schemes using [3+3]-cycloaddition reaction of 2-azidoacrylic acid derivatives with pyrrolphenylketone as a key stage. The anxiolytic activity of new substances has been established using open field test with flash. Results: Several synthesized N,1-diphenylpyrrolo[1,2-a]pyrazine-3-carboxamides derivatives significantly increased the total motor activity of Balb/c mice compared to the control. The structure-activity relationship was investigated. The most effective compound was found to be GML-11 (N-benzyl-N,1-diphenylpyrrolo[1,2-a]pyrazine-3-carboxamide), which had anxiolytic action in the dose range from 0.001 to 0.100 mg/kg (Balb/c, i.p.). This compound is two orders of magnitude higher in dose activity than all other pyrrolo[1,2-a]pyrazine TSPO ligands. Conclusion: Molecular modelling methods allowed us to create new TSPO ligands in the series of N,1-diphenylpyrrolo[1,2-a]pyrazine-3-carboxamides with high anxiolytic activity.

Screening of anxiolytic properties and analysis of structure-activity relationship of new derivatives of 6-(4-methoxy)-7H-[1,2,4]triazolo[3,4-a][2,3]benzodiazepine under the code RD

Introduction: Searching for new compounds with anti-anxiety activity resulting from the combination of privileged scaffolds is a promising direction in medicinal chemistry and in the development of new drugs. Anxiolytic potential and cytotoxic properties of previously synthesized molecules, containing fragments of 2,3-benzodiazepine and 1,2,4-triazole – 6-(4-methoxyphenyl)-7H-[1,2,4]triazolo[3,4-A][2,3]benzodiazepines under the generic code RD were studied. Materials and methods: Screening for anxiolytic activity was performed on elevated plus maze (EPM) and open field (OF) test models. Structural and functional analysis of the anti-anxiety activity of the studied substances was carried out. A degree of muscle relaxant effect of the substances was assessed in the tests Grid, Wire, and Rotarod. A cytotoxicity study of RD compounds was carried out using an MTT assay on human hepatocellular carcinoma cells HepG2. Results and discussion: For a number of novel triazolo[3,4-a][2,3]benzodiazepine derivatives, a prominent anxiolytic activity was manifested in terms of EPM test. The results of OF test were consistent with the obtained data and confirmed the presence of the sought activity in the leading compounds. There was no significant effect on muscle tone for the compounds under study. It was observed that RD compounds possessed no cytotoxic properties and were safe for further studies in vivo. Conclusion: Among the new derivatives of 6-(4-methoxyphenyl)-7H-[1,2,4]triazolo[3,4-a][2,3]benzodiazepine under the code RD, substances (RD-4, 12, 13) with a high anxiolytic activity comparable to diazepam and tofisopam were found. The most promising compound is RD-4 due to its pronounced anxiolytic and low cytotoxic properties.

Investigation of Potential Antioxidant, Thrombolytic and Neuropharmacological Activities of Homalomena aromatica Leaves Using Experimental and In Silico Approaches

The leaves of Homalomena aromatica are traditionally used in Bangladesh for the treatment of different chronic ailments. The purpose of this study was to explore in vitro antioxidant, thrombolytic activities, and in vivo neuropharmacological effects of methanolic extract of Homalomena aromatica (MEHA) leaves. Antioxidant activity of MEHA was assessed by a DPPH free radical scavenging assay and total phenolics content, total flavonoids content were also measured. The thrombolytic activity was determined by percentage of clot lysis and neuropharmacological activities by hole board, tail suspension, forced swimming and elevated plus maze tests. The results showed that the IC50 value of the extract against DPPH was 199.51 μg/mL. Quantitative analysis displayed higher contents of phenolics and flavonoids (147.71 mg gallic acid equivalent/g & 66.65 mg quercetin equivalent/g dried extract, respectively). The extract also showed a significant clot lysis (33.31%) activity. In case of anxiolytic activity, the elevate plus maze (EPM) test demonstrated an increase in time spent in open arms, and in case of hole board test, the number of head dipping was also significantly increased (p < 0.05). All the test compared with control (1% Tween in water) and standard (diazepam 1 mg/kg), significant dose (200 & 400 mg/kg) dependent anxiolytic activity was found. In antidepressant activity, there was a significant decrease in period of immobility in both test models (tail suspension and forced swimming) (p < 0.05). Moreover, 13 compounds were identified as bioactive, showed good binding affinities to xanthine oxidoreductase, tissue plasminogen activator receptor, potassium channel receptor, human serotonin receptor targets in molecular docking experiments. Furthermore, ADME/T analysis revealed their drug-likeness, likely pharmacological actions and non-toxic upon consumption. Taken together, our finding support the traditional medicinal use of this plant, which may provide a potential source for future drug discovery.

Coumarins from Seseli devenyense Simonk.: Isolation by Liquid–Liquid Chromatography and Potential Anxiolytic Activity Using an In Vivo Zebrafish Larvae Model

Different types of anxiety disorders have become the number one mental health issue in developed countries. The search for new, safer and effective drug-like molecules among naturally derived substances faces two difficulties: an efficient method of isolation compounds with a high-purity and high-throughput animal model for activity assay. Thus, the aim of the present study was to isolate by liquid–liquid chromatography high-purity rare coumarins from the fruits of Seseli devenyense Simonk. and evaluate their anxiolytic effect (defined as reversed thimotaxis) using a 5-days post-fertilization (dpf) Danio rerio larvae model. Liquid–liquid chromatography enabled the isolation of one simple hydroxycoumarin (devenyol) and four pyranocoumarins (cis-khellactone, d-laserpitin, isolaserpitin and octanoyllomatin). The anxiolytic effect was defined as a decrease in the time spent in the boundaries of the living space (also described as reversed thigmotaxis). Our results show that all isolated courmarins exerted a significant influence on the anxiety behavior (anxiolytic activity) in the zebrafish larvae model. According to our knowledge, this is the first report of anxiolytic activity of pyranocoumarins and devenyol.