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Ayahuasca is a psychoactive beverage that contains the psychoactive compound N,N-dimethyltryptamine and β-carboline alkaloids. This study aims at determining in vitro the bioavailability and bioaccessibility of the main compounds present in decoctions of four individual plants, in a commercial mixture and in four mixtures of two individual plants used in the preparation of Ayahuasca. The samples were subjected to an in vitro digestion process, and the Caco-2 cell line was used as an absorption model. The integrity and permeability of the cell monolayer were evaluated, as well as the cytotoxicity of the extracts. After digestion and cell incubation, the compounds absorbed by the cell monolayer were quantified by high-performance liquid chromatography coupled to a diode array detector. The results showed that compounds such as N,N-dimethyltryptamine, Harmine, Harmaline, Harmol, Harmalol and Tetrahydroharmine were released from the matrix during the in vitro digestion process, becoming bioaccessible. Similarly, some of these compounds, after being incubated with the cell monolayer, were absorbed, becoming bioavailable. The extracts did not show cytotoxicity after cell incubation, and the integrity and permeability of the cell monolayer were not compromised.

Peganum harmala seed extract has been frequently reported to possess antibacterial potential through in vivo studies. P. harmala L. (Zygophyllaceae) is one of the most famous medicinal plants used in traditional medicine of Iraq. The harmaline, harmalol and harmine exerted many pharmacodynemic effects on the central nervous system: stimulation and depression depending on the dose. P. harmala indicates a great variety of pharmacological activities such as antimicrobial, antitumor, antinociceptive and monoamine oxidase (MNO) inhibitory activities. The most important components of P. harmala seeds are harmaline, vasicinone and deoxyrsinone. The antibacterial effect of P. harmala was studied. The antibacterial activity of aqueous extracts was determined by agar well diffusion method. It inhibited the growth of Escherichia coli and Staphylococcus aureus. All animals injected with 100 mg/kg b. w of aqueous extract of P. harmala show myorelaxation or incoordination; so the animals dropped down from the wire 3 consecutive times i n 60 s. Aqueous extract of P. harmala also induced muscle relaxation and prolonged the sleeping time induced by pentoparpitol. These data suggest that P. harmala extract could inhibit the growth of S. aureus and E. coli strain in vitro and this activity may contribute to its chemopreventive effect. Key words: Antibacterial, muscle relaxant, hypnotic, Peganum harmala.

Traditional corrosion inhibitors have been frequently used for metal protection against strong acids. However, most of them are hazardous/toxic compounds with high cost. Therefore, due to the strict environmentally friendly regulation the green and low-cost compounds from renewable sources like plant extracts have gained high consideration in recent applications. In the present study, the aqueous Peganum harmala seed extract, commonly called Esfand, a cost-effective and green source of amine-rich molecules like harmaline, harmalol, harmane, harmol, vasicine and vasicinone, was selected to inhibit mild steel corrosion in 1 mol·L−1 HCl media. The Peganum harmala seed extract composition was investigated by FT-IR and UV–Visible analysis. The Peganum harmala seed extract corrosion inhibition behavior on mild steel in HCl solution was examined through electrochemical methods (EIS and polarization), surface analyses (AFM, SEM and contact angle) and theoretical studies (MD, DFT and MC). The presence of many electron donor atoms based on nitrogen and oxygen in the molecules of Peganum harmala seed extract is responsible for its effective adsorption behavior on metal surface through chelation with iron atoms. Experimental exploration declared that with the increase in Peganum harmala seed extract concentration and time of immersion the inhibition power increased and reached the high value of 95%. Peganum harmala seed extract behaved as a mixed type inhibitor and properly obeyed an adsorption isotherm of Freundlich. Results of AFM and SEM analyses clearly proved that Peganum harmala molecules adsorption on the metallic surface considerably reduced its dissolution rate and resulted in clean and smooth surface. The use of water instead of ethanol for extraction of major organic compounds of Peganum harmala seed, being non-toxic and cost-effective and exhibiting inhibition efficiency much greater than other green inhibitors (which are almost in the range of 80–90%), propose the Peganum harmala seed as a unique green corrosion inhibitor. Furthermore, the detailed theoretical observations elucidated from molecular simulations and density functional theory affirmed the Peganum harmala compounds adhesion via reactive sites.

The work highlighted interaction of harmalol, harmaline and harmine with human serum albumin by biophysical and biochemical assays. Presence of serum protein in the media negatively affects the cyt...

In a continuing effort to find new cytotoxic, antitumor, and less toxic agents from β-carbolines derivatives, using experimental and computational studies, five β-carboline alkaloids were tested in vitro for their cell growth inhibitory activity using Lepidium sativum phytotest. Then, molecular docking and correlation analysis between activity and structure study was performed to identify the interaction mode between these compounds and the colchicine binding site of tubulin. The experimental results revealed that harmaline and harmalol represent the most active compounds with IC50 of 134.15 μg/mL and 239.43 μg/mL, respectively, lower than colchicine which is 248 μg/mL. The correlation analysis between activity and structure indicates that the hydroxy group and the partial hydrogenation of the pyridyl ring increase activity. Indeed, molecular docking analysis revealed that harmaline and harmalol have low binding energy and could interact, in particular through conventional hydrogen bonds and van der Waals interactions with the colchicine binding site of tubulin, indicating that harmaline and harmalol could have an antimitotic effect. Moreover, in silico analysis of ADME/Tox properties reveals that these molecules possess promising pharmacokinetic properties in terms of intestinal adsorption, volume of distribution, blood-brain barrier permeability and toxicity. The results obtained were used to design new harmaline derivative molecule, with promising results after a preliminary in silico evaluation. In summary, our results indicate that the β-carboline alkaloids studied have an in vitro and in silico antimitotic activity and their pharmacokinetic properties are promising and should be confirmed by an in vivo study.

Parkinson’s Disease (PD) is a neurodegenerative disease that causes damage to the cognitive and motor system due to the death of dopaminergic neurons, which are responsible for the synthesis of the neurotransmitter dopamine. The study aimed to compare the monoamine oxidase B (MAO-B) inhibitory activity of natural molecules described in the literature with Selegiline, as potential drugs for the treatment of PD through molecular modeling, molecular docking and prediction of ADME/Tox properties. Thus, it was found the structure of the four natural molecules, Amburoside A, Harman, Harmaline and Harmalol, showed antiparkinsonian biological activity. Maps Electrostatic Potential showed similar regions between the molecules, except for Amburoside A, and Harmaline had a greater similarity in the positive potential with Selegiline. Molecular docking demonstrated that the studied molecules interact with 4–6 amino acids from the active site of the MAO-B enzyme, indicating that it has an inhibitory action on the enzyme, through hydrogen bonding and hydrophobic interactions. For ADME property predictions, most of the molecules showed good human oral absorption, all showed average permeability in Caco-2 cells, most showed average permeability in MDCK cells, showed low binding to plasma proteins, and for permeability in the blood-brain barrier, they were between good and medium. Overall, Harmaline has more properties similar to Selegiline. For toxicological properties, all molecules including Selegiline showed a positive result for the possibility of mutagenicity, whereas for the parameter of carcinogenicity in rats only the molecules Harmaline and Harmalol were positive, but no molecule was positive for carcinogenicity in mice. Therefore, the molecule that presented the best results was Harmaline, opening perspectives for the execution of in vitro studies.

In order to find new, less toxic and eco-friendly pest control agents, natural products are considered to be alternative options of potentially active compounds. In this context, the aim of the present study was the isolation and structural elucidation of the β-carboline and quinazoline alkaloids of Peganum harmala L. seeds. Silver nanoparticles (AgNPs) were prepared using alkaloids through a green synthesis procedure. Harmala alkaloids and their AgNPs showed considerable insecticidal and growth inhibitory activities against khapra beetle, Trogoderma granarium (Everts) (Coleoptera: Dermestidae). On a toxicity bioassay using treated filter papers, the total harmala alkaloids (THAs) were the most toxic followed by harmaline, harmine and harmalol, where LC50’s ranged between (30.6–61.7 µg/cm2) and (24.4–46.1 µg/cm2) 24 h post-treatment against larvae and adults, respectively. The two quinazolines, vasicine and vasicinone, showed moderate to weak activities. At 48 h post-exposure, efficacy of all alkaloids was increased. When tested as AgNPs (size 22.5–66.2 nm diameter), toxicity of alkaloids, especially the β-carbolines, was strongly increased, where LC50 values ranged between (4.7–11.4 µg/cm2) and (4.1–10.2 µg/cm2) 48 h post-treatment against larvae and adults, respectively. Feeding the 2nd instar larvae AgNPs-treated grains at sublethal concentrations led to significant drawbacks on the normal growth and development of the insect. These effects appeared as a high percentage of malformed larvae and pupae, a prolonged life span and a significant reduction in adult emergence. Results suggest the potential of using harmala alkaloids and their AgNPs as a natural approach for controlling T. granarium.

10 pags., 7 figs., 4 tabs., Syrian Rue, obtained from the seeds of the Peganum harmala plant, is a reddish colorant especially popular for dyeing carpets and wool fabrics in Western Asia and traditionally used as an alternative source for the madder-derived Turkey Red dye. The main constituents of the Syrian Rue dye are harmane, harmine, harmaline and harmalol, which belong to a group of substances collectively known as harmala alkaloids. These compounds are also called β-carboline alkaloids, as they display very similar molecular structures that consist of either a pyridine ring or a nitrogen-containing six-atom ring with two conjugated double bonds, each fused to an indole skeleton. In the present study, density functional theory was employed to perform a vibrational analysis of the four constituents of Syrian Rue by means of the Gaussian package. First, the molecular geometry of each compound was optimized and its Raman spectrum was calculated. Then, all the observed Raman bands were assigned to specific vibrational normal modes of the corresponding molecule. As a final step, the FT-Raman and surface-enhanced Raman spectroscopy (SERS) spectra of the four structurally related alkaloids were examined and compared, with the ultimate aim to study the adsorption mechanism of these molecules on silver nanoparticles used as SERS substrates., This research was supported by CSIC (Spanish National Research Council) through I-link+ Internalization Program (project i-link1148) and Ministerio de Economia, Industria y Competitividad de España (project FIS2017-84318-R). We are also indebted to the National Science Foundation (CHE1041832)

β-carboline alkaloids of the harmala group (HAlks)—a family of compounds with pharmacologic effects—can be found at trace levels ( −1 algae) in the edible invasive algae Undaria pinnatifida , known commonly as wakame. In this study, we present a simple and sensitive method to detect and quantify at low parts-per-trillion levels the six HAlks more frequently found in those plants. The method is based on on-line solid phase extraction capillary electrophoresis mass spectrometry using a C 18 sorbent. First, the methodology was optimized and validated with standard solutions through the use of ultraviolet (UV) and mass spectrometry (MS) detection. Second, the optimized method for MS detection was applied to an analysis of the HAlks in U. pinnatifida extracts. The method achieved limits of detection between 2 and 77 pg mL −1 for standards, producing an analyte preconcentration of about 1000-times in comparison to CE-MS. Some matrix effects were observed for the complex wakame extracts, especially for the most polar HAlks (harmol and harmalol), which bear aromatic hydroxyl groups. Harmine, harmaline, and norharmane were not detected in the algal extracts, whereas harmane was found at 70 pg mL −1 (70 ng kg −1 dry algae). The results underscored that C 18 -SPE-CE-MS may be considered as a powerful method to detect trace levels of alkaloids and other bioactive small molecules in complex plant extracts.

Objective To investigate the antibacterial effect of Peganum harmala ( P. harmala ) extracts against urinary tract infection pathogens isolated from infected patients. Methods Agar diffusion technique was used for detecting the antibacterial activity. The minimum inhibitory concentration was tested by serial dilution methods. Alkaloids extract of seeds was fractioned using thin layer chromatography. Results Phytochemical screening in the leaves of P. harmala showed the presence of flavonoids, alkaloids, saponins, tannins, glycosides, terpenoids and steroids and the absence of anthraquinons, whereas the results in the seeds showed the absence of flavonoids and the presence of alkaloids, saponins, tannins, glycosides, anthraquinons, terpenoids and steroids. The results of thin layer chromatography revealed that the alkaloids profiles of the seeds extracts of P. harmala are different. The major alkaloids detected and quantified from the intensity of their fluorescence were harmine, harmaline, harmalol and harmol. The methanolic extract of the plant was effective against the four microorganism tested. It was observed that the minimum inhibitory concentration and minimum bactericidal concentration of flavonoids extracted from the leaves and seeds of P. harmala were bactericidal to Staphylococcus aureus . Conclusions Results of this study suggest that the flavonoids extract of P. harmala may be useful to treat uropathogenic bacteria.

Harmalol administration caused remarkable reduction in proliferation of HepG2 cells with GI50 of 14.2 μM, without showing much cytotoxicity in embryonic liver cell line, WRL-68. Data from circular dichroism (CD) and differential scanning calorimetric (DSC) analysis of harmalol-CT DNA complex shows conformational changes with prominent CD perturbation and stabilization of CT DNA by 8 °C. Binding constant and stoichiometry was calculated using the above biophysical techniques. The Scatchard plot constructed from CD data showed cooperative binding, from which the cooperative binding affinity (K’ω) of 4.65 ± 0.7 × 105 M−1, and n value of 4.16 were deduced. The binding parameter obtained from DSC melting data was in good agreement with the above CD data. Furthermore, dose dependent apoptotic induction ability of harmalol was studied in HepG2 cells using different biochemical assays. Generation of ROS, DNA damage, changes in cellular external and ultramorphology, alteration of membrane, formation of comet tail, decreased mitochondrial membrane potential and a significant increase in Sub Go/G1 population made the cancer cell, HepG2, prone to apoptosis. Up regulation of p53 and caspase 3 further indicated the apoptotic role of harmalol.

The β-carboline alkaloids of the harmala (HAlks) group are compounds widely spread in many natural sources, but found at relatively high levels in some specific plants like Peganum harmala (Syrian rue) or Banisteriopsis caapi. HAlks are a reversible Mono Amino Oxidase type A Inhibitor (MAOI) and, as a consequence, these plants or their extracts can be used to produce psychotropic effects when are combined with psychotropic drugs based on amino groups. Since the occurrence and the levels of the HAlks in natural sources are subject to significant variability, more widespread use is not clinical but recreational or ritual, for example B. caapi is a known part of the Ayahuasca ritual mixture. The lack of simple methods to control the variable levels of these compounds in natural sources restricts the possibilities to dose in strict quantities and, as a consequence, limits its use with pharmacological or clinical purposes. In this work, we present a fast, simple, and robust method of quantifying simultaneously the six HAlks more frequently found in plants, i.e., harmine, harmaline, harmol, harmalol, harmane, and norharmane, by capillary electrophoresis instruments equipped with the more common detector UV. The method is applied to analyze these HAlks in P. Harmala seeds infusion which is a frequent intake form for these HAlks. The method is validated in three different instruments in order to evaluate the transferability and to compare the performances between them. In this case, harmaline, harmine, and harmol were found in the infusion samples. Copyright © 2016 John Wiley & Sons, Ltd.

An extraction method based on solid-phase extraction was developed to determine the major alkaloid components, N,N-dimethyltryptamine, harmine, harmaline, harmalol, and tetrahydroharmine, in ayahuasca using ultra-performance liquid chromatography-tandem mass spectrometry. Coconut charcoal and menthone-thiosemicarbazone polymer were tested as new adsorbent materials. Experiments carried out at levels of 1, 5, 10, 50, 100, and 250 µg mL-1 resulted in recoveries in the ranges 45.3-115.7% for menthone-thiosemicarbazone and 40.6-116.2% for coconut charcoal. The detection and quantification limits ranged from 0.003 to 0.016 µg mL-1 and from 0.009 to 0.043 µg mL-1, respectively. The method developed was linear over the tested range (0.05-1.0 µg mL-1), with correlation coefficients ranging from 0.9969 to 0.9998. The technique was applied for analysis of the compounds in an ayahuasca sample collected from a religious group located in the city of Fortaleza, obtaining concentrations in the range from 0.3 to 19.3 mg mL-1.

Poor water solubility of pharmacologically active ingredients causes to incomplete absorption through the gastrointestinal tract. Increasing surface area by micronization has been performed to overcome these problems. In this study, the first step entailed an optimization of supercritical fluid extraction from the capsules of Peganum harmala L. As the second step, micronization process was accomplished using a new technique based on the expansion of CO2 supercritical solvent and collected precipitates were characterized by Field Emission Scanning Electron Microscopy (FESEM). The herbal extract was qualified using HPLC- mass spectrometry and Harmaline, Harmine and Harmalol were identified as the main β-carboline alkaloids. Effect of different supercritical parameters on the extraction yield and particle size were investigated. As a result, particle size decreased as the precipitation time decreased, whereas temperature and loaded volume did not significantly affect the morphology. Optimum micronization conditions were determined as: Volume of loaded extract (50 μL), oven temperature (40 °C), equilibrium and precipitation pressure (350 and 100 bar), equilibration and precipitation time (10 min). This supercritical process is proposed as a green technique for extraction and preparation of micronized β-Carbolines and nanoparticles with an average diameter ranging from 7 to 100 nm were successfully formed.

Ethnopharmacological relevance The seeds of Peganum harmala Linn, in which the most abundant active compounds are harmaline and harmine, have been widely used as a traditional medicine in various countries to treat a broad spectrum of diseases including asthma, cough, depression, Parkinson's and Alzheimer's diseases. However, few studies on long-term or subchronic toxicity of seeds of P. harmala were reported after overdose. Aim of the study To investigate the subchronic toxicity and concomitant toxicokinetics of total alkaloid extracts from seeds of P. harmala (TAEP) after oral administration for four weeks in rats. Materials and methods The subchronic toxicity and concomitant toxicokinetics of TAEP were evaluated after 28-day oral administration in rats at daily dose levels of 15, 45, and 150 mg/kg. The signs of toxicity and mortality were monitored and recorded daily. The body weight and average food consumption were measured weekly. The analyses of hematology, biochemistry, urine, relative organ weights and histopathology were conducted at the termination of treatment and recovery phase. For concomitant toxicokinetics study, the plasma toxicokinetic parameters, tissue distribution, and excretion of predominant ingredients harmaline and harmine in TAEP and metabolites harmalol and harmol were tested. Results Following initial repeated exposure to high-dose (150 mg/kg/day) of TAEP excitotoxic reaction, such as tremor, was observed, but tolerated on the fourth day after multiple dosing. The significant alterations in blood glucose and lipid metabolism in liver were observed, but recovered after four weeks of drug withdrawal. The no-observed-adverse-effect level (NOAEL) of TAEP was considered to be 45 mg/kg/day under the present study conditions. There were no significant gender differences in most indexes of subchronic toxicity throughout the experimental period with the exception of food consumption and body weight. In concomitant toxicokinetics study, the alterations of dynamic characteristic for harmaline, harmine and metabolite harmol after multiple oral administration at three doses had been observed. Harmaline, harmine and metabolites harmalol and harmol were widely distributed in organs and there was no accumulation in the tissues examined. The reduction of harmaline and metabolite harmalol in brain after multiple dosing at dose of 150 mg/kg might be closely related to the tremor tolerance. The main excretory pathway for metabolites harmalol and harmol was urinary excretion via kidney. Conclusions The results revealed that TAEP at doses of 15 and 45 mg/kg/day in rats might be safe. Excitotoxic reaction such as tremor occurred initially at dose of 150 mg/kg/day, however, the toxicity was tolerant and reversible. In addition, harmaline and harmine in TAEP had a quick absorption into blood and metabolized to harmalol and harmol, and there was no drug accumulation in the detected tissues. Further studies should be investigated to clarify the mechanisms of tremor tolerance and neurotoxicity of TAEP.

A simple method based on micellar liquid chromatography (MLC) was developed to enable the simultaneous determination of norharmane, harmane, harmol, harmalol, harmine, and harmaline in phytotherapic samples of Passiflora incarnata L., Passiflora alata Dryander and urine with virtually no sample preparation. Chromatographic separation was performed under 30 min using a C18 HPLC column and isocratic elution with an aqueous micellar mobile phase (phosphate buffer pH 8.0 containing 220 mmol L−1 of sodium dodecyl sulfate and 0.5% of triethylamine) containing only 3% (v/v) of acetonitrile. Fluorescence detection allowed limits of quantification below 4.5 ng g−1. Recoveries in controlled samples were close to 100% and the sample analysis enabled the quantification of harmol as the principal β-carboline found in the phytotherapic samples, whereas another three were detected. In urine samples, the determination of harmol was made after 8 hours of drug ingestion. Both repeatability and intermediary precision, for a...

This is the first report on the development and validation of high-performance thin-layer chromatography (HPTLC) method for simultaneous analysis of five antipsychotic and medicinally important β-carboline alkaloids (βCAs), namely, harmalol, harmaline, harmine, harmane and norharmane. These βCAs occurs in both plant and animal system including human being. In the present investigation, their best separation was achieved using an optimized mobile phase, chloroform: methanol: glacial acetic acid (7.8:2.2:0.2, v/v/v) on aluminum TLC plates precoated with silica gel 60 F254. The quantification was performed by densitometric scanning in fluorescence mode at 366 nm. The calibration curves were drawn using linear regression, plotted over the range 25-250 ng band-1 of standard βCAs with correlation coefficient (R2) between 0.97 and 0.992. Accuracy in terms of recovery (83.95-112.40%), repeatability of application (0.61-2.42%), repeatability of measurement (1.94-3.05%) and intermediate precision (0.62-11.16%) of developed method were simultaneously determined. The limit of detection and limit of quantification were between 4.95-6.59 and 16.50-21.93 ng band-1, respectively. The method was validated according to ICH guidelines and was simple, cost-effective, precise, sensitive and specific for the determination of βCAs in herbs, Fagonia schweinfurthii, Peganum harmala and Tribulus terrestris. The developed HPTLC method would have importance in forensic and industrial chromatographic analysis and fingerprinting of various herbs and drug formulations containing βCAs.

β-carbolines (βCs) are alkaloids widely distributed in nature that have demonstrated antimicrobial properties. Here, we tested in vitro six βCs against Penicillium digitatum and Botrytis cinerea, causal agents of postharvest diseases on fruit and vegetables. Full aromatic βCs (harmine, harmol, norharmane and harmane) exhibited a marked inhibitory effect on conidia germination at concentrations between 0.5 and 1 mM, while dihydro-βCs (harmalina and harmalol) only caused germination delay. Harmol showed the highest inhibitory effect on both fungal pathogens. After 24 h of exposure to 1 mM harmol, conidia revealed a severe cellular damage, exhibiting disorganized cytoplasm and thickened cell wall. Harmol antimicrobial effect was fungicidal on B. cinerea, while it was fungistatic on P. digitatum. Conidia membrane permeabilization was detected in treatments with harmol at sub-inhibitory and inhibitory concentrations, for both pathogens. In addition, residual infectivity of P. digitatum on lemons and B. cinerea on blueberries was significantly reduced after exposure to this alkaloid. It also inhibited mycelial growth, preventing sporulation at the highest concentration tested. These results indicate that harmol might be a promising candidate as a new antifungal molecule to control causal agents of fruit diseases. Fil: Olmedo, Gabriela María. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto Superior de Investigaciones Biológicas. Universidad Nacional de Tucumán. Instituto Superior de Investigaciones Biológicas; Argentina Fil: Cerioni, Luciana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto Superior de Investigaciones Biológicas. Universidad Nacional de Tucumán. Instituto Superior de Investigaciones Biológicas; Argentina Fil: Gonzalez, Maria Micaela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina Fil: Cabrerizo, Franco Martín. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina Fil: Rapisarda, Viviana Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto Superior de Investigaciones Biológicas. Universidad Nacional de Tucumán. Instituto Superior de Investigaciones Biológicas; Argentina Fil: Volentini, Sabrina Inès. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto Superior de Investigaciones Biológicas. Universidad Nacional de Tucumán. Instituto Superior de Investigaciones Biológicas; Argentina

Herbal remedies for human brain disorders are much preferred over synthetic drugs because of various side effects of synthetic drugs ranging from sleep disorders to withdrawal syndromes. Passiflora incarnata Linn (Passifloraceae) has been widely used in traditional medicine in West India, Mexico, the Netherlands, and South America. It has been used as an anxiolytic and sedative-hypnotic since ancient times. The main constituents of leaves of P. incarnata are flavonoid and indole alkaloids based on β -carboline ring system viz., harman, harmine, harmalol and harmaline. Harmine and harmaline alkaloids are reported to be effective antiparkinsonian compounds. The objective of present investigation was to evaluate antiparkinsonian activity and antioxidant activity of butanol extract of P. incarnata leaves (BEPI). Antiparkinsonian activity of BEPI was studied using haloperidol induced catalepsy and tacrine induced jaw movements. The antioxidant activity of BEPI was studied using DPPH scavenging assay and H2O2 scavenging assay. The results of present study has shown that butanol extract of P. incarnata leaves (BEPI) (150 mg kg−1 and 300 mg kg−1, i.p.) significantly (p ≤ 0.001) reduced the duration of haloperidol induced catalepsy as well as tacrine induced jaw movements. BEPI has also shown significant antioxidant effect. The result obtained in current study indicates that BEPI posses antiparkinsonian as well as antioxidant activity which may be useful in symptomatic relief of PD and to prevent progression of PD.

Harmalol exhibits pH dependent structural equilibrium between protonated and deprotonated forms with a p K a of 7.8 as revealed from spectroscopic titration. The compound exists as protonated (structure I) and deprotonated (structure II) form in the pH range 1–7 and 9–12, respectively. The interaction of structure I and II to calf thymus DNA has been studied by different spectroscopic and calorimetric techniques in buffer of pH 6.8 and 9.2, respectively. The results show that structure I bind strongly to DNA showing a cooperative mode with a binding constant of 4.5 × 10 5 M −1 and a stoichiometry of 4.8 nucleotide phosphates. The alkaloid stabilized the DNA by 8 °C, the binding shows 40% quenching of fluorescence intensity, perturbation in circular dichroism spectra and enthalpy driven exothermic binding with a large hydrophobic contribution to the binding free energy. Furthermore, the alkaloid shows a prominent change of specific viscosity with sonicated linear DNA and unwinding–rewinding of covalently closed pUC 18 DNA, revealing intercalative binding. The deprotonated structure (structure II), on the other hand, in the presence of large amount of DNA concentration, converts back to a structure I-DNA complexation. This transition has been presumably induced by the polyanionic phosphate backbone of DNA at high concentration.

Peganum harmala (L) is a perennial plant which is native of eastern Iranian and west of India but also found in different regions of western USA. A number of β-carboline compounds with therapeutic importance and different pharmacological effects, are present in this plant. Among other alkaloids, such as, harmine, harmalol and vasicine, isolated from various parts of the plant, harmaline is considered as most valuable with reference to its medicinal importance. Harmaline has been extensively studied in last decade and known to exert multiple pharmacological effects including antileishmanial, antimicrobial, antiplatelet, antiplasmodial, antitumoral, hypothermic and vasorelaxant activity. The proposed work is intended to highlight the recent pharmacological aspects of β-carboline alkaloid "harmaline".

Harmaline and harmine accounted for more than 70% in composition in extracts of P. harmala. More attention, however, should be paid to the other alkaloids which would be favorable or unfavorable to the efficacy and safety of the products. It was necessary to determine these trace alkaloids in the extracts; thereafter, most of them have been characterized. Diglycoside vasicine, vasicine, vasicinone, harmalol, harmol, tetrahydroharmine, 8-hydroxy-harmine, ruine, harmaline, and harmine were separated and identified with reference substances and characteristic MS spectra in extracts by liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) and high-performance liquid chromatography (HPLC). Three trace alkaloids, vasicine, harmalol, and harmol were determined using the developed chromatographic separation method subsequently. The average contents of vasicine, harmalol, and harmol in extracts of ten batches were 2.53 ± 0.73, 0.54 ± 0.19, and 0.077 ± 0.03%, respectively. The total content of the three alkaloids was 3.23 ± 0.90% (from 1.81 to 4.48%). For rough estimation of all the relative alkaloids except of harmaline and harmine, the average total areas of all peaks in extracts varied from 4.35 to 26.64% detected at 220, 254, 265, 280, and 380 nm, respectively. The results indicated that area normalization method was powerless for the quality evaluation for traditional herb medicine consisting of numerous compounds with highly differential features. It might be concluded that LC-MS or HPLC could be utilized as a qualitative and quantitative analytical method for quality control of the extracts from seeds of P. harmala L.

Background Harmalol, a beta carboline alkaloid, shows remarkable importance in the contemporary biomedical research and drug discovery programs. With time, there is emerging interest in search for better anti-cancer drugs of plant origin with high activity and lower toxicity. Most of the chemotherapeutic agents due to their non-specific target and toxicity on active healthy cells, use is often restricted, necessitating search for newer drugs having greater potentiality. Objective The review highlighted the interaction of harmalol with nucleic acids of different motifs as sole target biomolecules and in vitro cytotoxicity of the alkaloid in human cancer cell lines with special emphasis on its apoptotic induction ability. Methods Binding study and in vitro cytotoxicity was performed using several biophysical techniques and biochemical assays, respectively. Results Data from competition dialysis, UV and fluorescence spectroscopic analysis, circular dichroism, viscometry and isothermal calorimetry shows binding and interaction of harmalol with several natural and synthetic nucleic acids, both DNA and RNA, of different motifs. Furthermore, apoptotic hallmarks like internucleosomal DNA fragmentation, membrane blebbing, cell shrinkage, chromatin condensation, change of mitochondrial membrane potential, comet tail formation and ROS (reactive oxygen species) dependent cytotoxicity being analyzed in the harmalol treated cancer cells. Conclusion These results stating the therapeutic role of harmalol, will lead to the interesting knowledge on the cytotoxicity, mode, mechanism, specificity of binding and correlation between structural aspects and energetics enabling a complete set of guidelines for design of new drugs.

Ayahuasca is a potent hallucinogenic beverage prepared from Banisteriopsis caapi in combination with other psychoactive plants. N,N-dimethyltryptamine, tryptamine, harmine, harmaline, harmalol, and tetrahydroharmine were quantified in ayahuasca samples using a simple and low-cost method based on SPE and LC with UV diode-array detection. The experimental variables that affect the SPE method, such as type of solid phase and nature of solvent, were optimized. The method showed good linearity (r > 0.9902) and repeatability (RSD < 0.8%) for alkaloid compounds, with an LOD of 0.12 mg/L. The proposed method was used to analyze 20 samples from an ayahuasca cooking process from a religious group located in the municipality of Fortaleza, state of Ceará, Brazil. The results showed that concentrations of the target compounds ranged from 0.3 to 36.7 g/L for these samples.

Wild Syrian rue (Peganum harmala L. family Zygophyllaceae) is well-known in Iran and various parts of this plant including, its seeds, bark, and root have been used as folk medicine. Recent years of research has demonstrated different pharmacological and therapeutic effects of P. harmala and its active alkaloids, especially harmine and harmaline. Analytical studies on the chemical composition of the plant show that the most important constituents of this plant are beta-carboline alkaloids such as harmalol, harmaline, and harmine. Harmine is the most studied among these naturally occurring alkaloids. In addition to P. harmala (Syrian rue), these beta-carbolines are present in many other plants such as Banisteria caapi and are used for the treatment of different diseases. This article reviews the traditional uses and pharmacological effects of total extract and individual active alkaloids of P. harmala (Syrian rue).

Previous studies have shown an antimalarial effect of total alkaloids extracted from leaves of Guiera senegalensis from Mali in West Africa. We independently observed that the beta-carboline alkaloid harmine obtained from a natural product library screen inhibited Plasmodium falciparum heat shock protein 90 (PfHsp90) ATP-binding domain. In this study, we confirmed harmine-PfHsp90-specific affinity using surface plasmon resonance analysis (dissociation constant [ K d ] of 40 μM). In contrast, the related compound harmalol bound human Hsp90 (HsHsp90) ( K d of 224 μM) more tightly than PfHsp90 ( K d of 7,010 μM). Site-directed mutagenesis revealed that Arg98 in PfHsp90 is essential for harmine selectivity. In keeping with our model indicating that Hsp90 inhibition affords synergistic combinations with existing antimalarials, we demonstrated that harmine potentiates the effect of chloroquine and artemisinin in vitro and in the Plasmodium berghei mouse model. These findings have implications for the development of novel therapeutic combinations that are synergistic with existing antimalarials.

Ayahuasca is an Amazonian psychotropic plant tea obtained from Banisteriopsis caapi, which contains β-carboline alkaloids, chiefly harmine, harmaline and tetrahydroharmine. The tea usually incorporates the leaves of Psychotria viridis or Diplopterys cabrerana, which are rich in N,N-dimethyltryptamine (DMT), a psychedelic 5-HT2A/1A/2C agonist. The β-carbolines reversibly inhibit monoamine-oxidase (MAO), effectively preventing oxidative deamination of the orally labile DMT and allowing its absorption and access to the central nervous system. Despite increased use of the tea worldwide, the metabolism and excretion of DMT and the β-carbolines has not been studied systematically in humans following ingestion of ayahuasca. In the present work, we used an analytical method involving high performance liquid chromatography (HPLC)/electrospray ionization (ESI)/selected reaction monitoring (SRM)/tandem mass spectrometry(MS/MS) to characterize the metabolism and disposition of ayahuasca alkaloids in humans. Twenty-four-hour urine samples were obtained from 10 healthy male volunteers following administration of an oral dose of encapsulated freeze-dried ayahuasca (1.0 mg DMT/kg body weight). Results showed that less than 1% of the administered DMT dose was excreted unchanged. Around 50% was recovered as indole-3-acetic acid but also as DMT-N-oxide (10%) and other MAO-independent compounds. Recovery of DMT plus metabolites reached 68%. Harmol, harmalol, and tetrahydroharmol conjugates were abundant in urine. However, recoveries of each harmala alkaloid plus its O-demethylated metabolite varied greatly between 9 and 65%. The present results show the existence in humans of alternative metabolic routes for DMT other than biotransformation by MAO. Also that O-demethylation plus conjugation is an important but probably not the only metabolic route for the harmala alkaloids in humans. Copyright © 2012 John Wiley & Sons, Ltd.

Harmala alkaloids are pharmaceutically active molecules that can be found in various plants. These alkaloids are fluorescent molecules in the range of 300-700nm. Coincidently, bees have a similar visible range of 300-600nm. This study takes these observations and interweaves them into a hypothesis: since bees use their sight to find flowers to pollinate, then these flowers contain harmala alkaloids that would be visible to bees. It can then be inferred harmala alkaloids attract bees. In other words, harmala alkaloids are functional components of plants. In order to determine harmala alkaloids content, standard solutions of harmine, harmaline, harmane, harmol, and harmalol will be compared with extractions from plant samples using high performance liquid chromatography and fluorescence.A variety of plants were chosen to represent three categories. The first is plants that are found to be insect pollinated, these include lemon balm (Melissa officinali), common rue (Ruta graveolens), meadow rue (Thalictrum aquilegifolium), hydrangea (Hydrangea arborescens), spirea (Spirea japonica), forget-me-not (Myosotis scorpioides), blue star grass (Sisyrinchium augustifolium), common rue (Ruta graveolens) and meadow rue (Thalictrum aquilegifolium). The second category represents wind pollinated plants, including sugar maple (Acer saccharum), white velvet (Tradescantia sillamontana), meadow rue (Thalictrum ichangense), rhoeo (Rhoeo spathacea). Finally, a control was also analyzed. The lady fern (Athyrium felix-femina) was chosen because it is not genetically related to the plants in categories one or two and is not insect or wind pollinated.Following chemical analysis, each of the insect pollinated plants was found to contain harmala alkaloids. The lady fern (Athyrium felix-femina) contained no harmala alkaloids, as well as the wind pollinated plants. Due to these results as well as a study of bee behavior, we were able to conclude that harmala alkaloids are present in plants that attract bees. This study both contributes to an understanding of factors involved in pollination and can be used as a guide for further investigation into a natural source of harmala alkaloids.

β-Carboline alkaloids are the main chemical constituents of the plant Peganum harmala, while they also could be formed endogenously and found in coffee, alcoholic beverages and tobacco. Considering the fact that the possibility of herb–drug interactions has recently received great attention worldwide, the aim of the current study was to assess the potential for the metabolism-based drug–drug interactions arising from five β-carboline alkaloids (harmine, harmaline, harmalol, harmol and harmane) from P. harmala in vitro. With microsome incubation assays and UPLC/HPLC methods, the inhibitions on human liver CYP3A4 and CYP2D6 enzymes by those β-carboline alkaloids were studied kinetically. Harmine, harmol and harmane exhibited noncompetitive inhibition on the activity of CYP3A4 with Ki values of 16.76, 5.13 and 1.66 μm, respectively. These β-carboline alkaloids were also found to be both substrates and inhibitors for CYP2D6. Harmaline, harmine and harmol showed typical competitive inhibition on the activity of CYP2D6 with Ki values of 20.69, 36.48 and 47.11 μm, respectively. The inhibition of the two major CYP enzymes by those β-carboline alkaloids suggested that changes in the pharmacokinetics of co-administered drugs were likely to have occurred. Therefore, caution should be exercised for possible drug interactions of medicinal plants containing those β-carboline alkaloids and CYP substrates. Copyright © 2011 John Wiley & Sons, Ltd.

Melanin synthesis is regulated by melanocyte specific enzymes and related transcription factors. β-carboline alkaloids including harmaline and harmalol are widely distributed in the environment including several plant families and alcoholic beverages. Presently, melanin content and tyrosinase activity were increased in melanoma cells by harmaline and harmalol in concentration- and time-dependent manners. Increased protein levels of tyrosinase, tyrosinase-related protein-1 (TRP-1), and TRP-2 were also evident. In addition, immunofluorescence and Western blot analyses revealed harmaline and harmalol increased cAMP response element binding protein phosphorylation and microphthalmia-associated transcription factor expression. In addition to studying the signaling that leads to melanogenesis, roles of the p38 MAPK pathways by the harmaline and harmalol were investigated. Harmaline and harmalol induced time-dependent phosphorylation of p38 MAPK. Harmaline and harmalol stimulated melanin synthesis and tyrosinase activity, as well as expression of tyrosinase and TRP-1 and TRP-2 indicating that these harmaline and harmalol induce melanogenesis through p38 MAPK signaling.

β-Carboline alkaloids are present in medicinal plants such as Peganum harmala L., which have been used as folk medicine in anticancer therapy. Recently, they have drawn attention because of their antitumor activities. Despite considerable interest and investigations on alkaloid-DNA complexes, reports on alkaloid-RNA interaction are very limited. This study is the first attempt to investigate the binding of β-carboline alkaloids (harmine, harmane, harmaline, harmalol, and tryptoline) with yeast RNA. The effect of alkaloid complexation on RNA aggregation and condensation was investigated in aqueous solution at physiological conditions, using constant RNA concentration (6.25 mM) and various alkaloid:polynucleotide (phosphate) ratios of 1:240, 1:160, 1:80, 1:40, 1:20, 1:10, 1:5, 1:2, and 1:1. Fourier transform infrared and UV-visible spectroscopic methods were used to determine the ligand-binding modes, the binding constants, and the stability of alkaloid-RNA complexes in aqueous solution. Spectroscopic evidence showed major binding of alkaloids to RNA with overall binding constants of K(harmine)-RNA = 2.95 × 10⁷ M⁻¹, K(harmane)-RNA = 5.62 × 10⁵ M⁻¹, K(harmaline)-RNA = 7.47 × 10⁵ M⁻¹, K(harmalol)-RNA = 4.32 × 10⁵ M⁻¹, and K(tryptoline)-RNA = 3.21 × 10⁵ M⁻¹. The affinity of alkaloids-RNA binding is in the order of harmine harmaline harmane harmalol tryptoline. No biopolymer secondary structural changes were observed upon alkaloid interaction and RNA remains in the A-family structure in these complexes.

Preliminary work on Passiflora alata leaves failed to detect harmane alkaloids using LC. The aim of this work was to investigate the production of harmane alkaloids through the cell culture of P. alata, inducing its precursor (L-tryptophan). The leaf explants presented satisfactory results after disinfection, and the callus formation was initiated in MS media with adequate quantities of phytohormones. Sixty days after inoculation, calli were inoculated in the optimized semi-solid MS media, with and without the addition of L-tryptophan (50, 100, 200 mg/L) and kept in standard conditions for 90 days. Calli were collected on days 6, 16, 26, 36, and 90, followed by acid-base extraction, and analysed by LC. The results showed an absence of harmane, harmin, harmol, harmalol, and harmaline. With L-tryptophan feeding, two peaks were detected, collected and analysed through positive mode electrospray [ESI(+)-MS] and sequential analysis in tandem ESI(+)-MS/MS. The spectra obtained were very similar, with a repetition of the more intense ions, and consecutive loss of 68 Da units, attributed to the heterocycle pyrazole. It appeared that this transformation was not related to any enzymatic pathway previously described for the plant from L-tryptophan, and the biosynthesis of β-carboline alkaloids in callus culture of P. alata were not observed in this work.As folhas de varias espécies de Passiflora são utilizadas como ansioliticas e sedativas. Passiflora alata Curtis, Passifloraceae consta em três edições da farmacopéia brasileira, porem não há muitos estudos sobre sua composição química. No passado, enfatizava-se a ação conjunta de alcalóides e flavonóides. Em trabalho anterior, não foi detectada a presença de alcalóides harmanicos através de CLAE. Assim, decidiu-se investigar a produção dos mesmos através de cultivo celular, introduzindo seu precursor metabólico L-triptofano. Os explantes foliares apresentaram resultados satisfatorios para germinação apos assepsia, e a formação de calo foi iniciada em meio MS com quantidades adequadas de fitohormonios, previamente determinadas. Sessenta dias após a inoculação os calos foram repicados para meio semi-solido com e sem L-triptofano (50, 100, 200 mg/L), mantidos por 90 dias em condições padrão. Amostras foram coletadas com 6, 16, 26, 36, e 90 dias, realizada extração acido-base e o extrato analisado por CLAE. Os resultados mostraram a ausência de harmana, harmina, harmol, harmalol e harmalina. Dois picos presentes nas amostras com L-triptofano foram coletados e analisados através de espectrometria de massas, electrospray modo positiva [ESI(+)-MS] e analise em tandem ESI(+)-MS/MS. Os espectros correspondentes foram similares, mostrando a perda consecutiva de 68 Da, atribuídos ao pirazol. Este fato aponta para uma transformação não enzimática, não relacionada a uma biossintese previamente descrita para alcalóides β-carbolínicos.

Beta-carboline alkaloids present in Peganum harmala (harmal) have recently drawn attention due to their antitumor activities. The mechanistic studies indicate that beta-carboline derivatives inhibit DNA topoisomerases and interfere with DNA synthesis. They interact with DNA via both groove binding and intercalative modes and cause major DNA structural changes. The aim of this study was to examine the interactions of five beta-carboline alkaloids (harmine, harmane, harmaline, harmalol and tryptoline) with calf-thymus DNA in aqueous solution at physiological conditions, using constant DNA concentration (6.25 mM) and various alkaloids/polynucleotide (phosphate) ratios of 1/240, 1/160, 1/80, 1/40, 1/20, 1/10, 1/5, 1/2 and 1/1. Fourier transform infrared (FTIR) and UV-visible spectroscopic methods were used to determine the ligand binding modes, the binding constants, and the stability of alkaloids-DNA complexes in aqueous solution. Spectroscopic evidence showed major binding of alkaloids to DNA with overall binding constants of K(harmine)-DNA=3.44x10(7) M(-1), K(harmane)-DNA=1.63x10(5) M(-1), K(harmaline)-DNA=3.82x10(5) M(-1), K(harmalol)-DNA=6.43x10(5) M(-1) and K(tryptoline)-DNA=1.11x10(5) M(-1). The affinity of alkaloids-DNA binding is in the order of harmine>harmalol>harmaline>harmane>tryptoline. No biopolymer secondary structural changes were observed upon alkaloid interaction and DNA remains in the B-family structure in these complexes.

The beta-carboline alkaloids are naturally existing plant substances. It is known that these alkaloids have a wide spectrum of neuropharmacological, psychopharmacological, and antitumor effects. Therefore, they have been traditionally used in oriental medicine for the treatment of various diseases including cancers and malaria. In this study, harmol and harmalol, which are beta-carboline alkaloids, were examined for their antitumor effect on human lung carcinoma cell lines, and structure-activity relationship was also investigated. H596, H226, and A549 cells were treated with harmol and harmalol, respectively. Apoptosis was induced by harmol only in H596 cells. In contrast, harmalol had negligible cytotoxicity in three cell lines. Harmol induced caspase-3, caspase-8, and caspase-9 activities and caspase-3 activities accompanied by cleavage of poly-(ADP-ribose)-polymerase. Furthermore, harmol treatment decreased the native Bid protein, and induced the release of cytochrome c from mitochondria to cytosol. The apoptosis induced by harmol was completely inhibited by caspase-8 inhibitor and partially inhibited by caspase-9 inhibitor. The antagonistic antibody ZB4 blocked Fas ligand-induced apoptosis, but had no effect on harmol-induced apoptosis. Harmol had no significant effect on the expression of Fas. In conclusion, our results showed that the harmol could cause apoptosis-inducing effects in human lung H596 cells through caspase-8-dependent pathway but independent of Fas/Fas ligand interaction.

Beta-carboline alkaloids including harmalol, harmaline, norharmane, harmol, harmine and harmane are important constituents of the medicinal plant, Perganum harmala L. (Zygophylaceae), which has been used in traditional medicine. In the present study, the antiplatelet activities of six beta-carboline alkaloid compounds were investigated in vitro. At a concentration of 200 microM, these compounds have no effect on arachidonic acid (AA)-, thrombin- and U46619 (a thromboxane A2 mimic)-stimulated platelet aggregation. On the contrary, it was revealed that collagen-induced platelet aggregation could be inhibited by these compounds with different potencies (harmane and harmine were most potent, harmol had medium potency, and harmol, norharmane, harmalol and harmaline had a weak, non significant effect), indicating a selective inhibition on collagen-mediated platelet activation. Consistently, further study revealed that collagen-mediated phospholipase (PL) Cgamma2 and protein tyrosine phosphorylation, cytosolic calcium mobilization and arachidonic acid liberation were completely inhibited by harmane and harmine in a concentration-dependent manner, while the other compounds were only partially or not effective at all. Taken together, these results indicate that three of these six beta-carboline alkaloids can selectively affect collagen-induced platelet aggregation with different potencies; in particular, harmane and harmine were most potent, and their antiplatelet activities may be mediated by inhibiting PLCgamma2 and protein tyrosine phosphorylation with sequential suppression of cytosolic calcium mobilization and arachidonic acid liberation, indicating that harmane and harmine have a potential to be developed as a novel agent for atherothrombotic diseases.

Chemical composition of the floral nectar of Peganum harmala, a herbaceous medicinal perennial of the family Zygophyllaceae, was analysed using high-performance liquid chromatography technique. The nectar sugar detection experiments resulted in 33.1, 39.8 and 27.4%, respectively, for fructose, glucose and sucrose, upon which the nectar was classified as hexose rich. In addition, 11 proteinaceous amino acids were recognised and quantified in the nectar. Concentration of the insects' favoured amino acid, prolin, was markedly high. Furthermore, among four detected alkaloids, harmalol and harmine as the two beta-carboline derivatives were identified. These findings may confer a better understanding upon outcrossing processes and favour the plant-pollinator relationships.