harmine and 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.

Topics: Alkaloids; Carbolines; Electrophoresis, Capillary; Harmaline; Harmine; Limit of Detection; Monoamine Oxidase Inhibitors; Peganum; Plant Extracts; Psychotropic Drugs; Seeds; Time Factors

Harmine (HAR) and harmaline (HAL) were metabolized by demethylation to form harmol (HOL) and harmalol (HAM) both in vivo and in vitro. It has been demonstrated tremendous value of HAR, HAL and their metabolites in the therapy of Alzheimer's disease. A rapid, selective and sensitive UPLC-ESI-MS/MS method was firstly developed and validated for the simultaneous determination of HAR, HAL, HOL, and HAM in beagle dog plasma with 9-aminoacridine as the internal standard (IS). After protein precipitation with acetonitrile, the analytes were separated within 4.5 min on an ACQUITY UPLC BEH C18 column with a gradient elution system composed of 0.1% formic acid and acetonitrile at a flow rate of 0.4 ml/min. Detection was performed using multiple reactions monitoring mode under a positive ionization condition. The calibration curves of four analytes showed good linearity (r(2)>0.9959) within the tested concentration ranges. The low limit of quantification for HAR, HAL, HOL, and HAM were all 1.00 ng/ml. The mean accuracy of the analytes was within the range of 94.56-112.23%, the R.S.D. values of intra-day and the inter-day precision were less than 6.26% and 7.51%, respectively. Matrix effects and extraction recoveries of the analytes from the beagle dog plasma were within the range of 94.48-105.77% and 89.07-101.44%, respectively. The validated method was successfully applied to a pharmacokinetic study of HAR, HAL, HOL, and HAM in beagle dogs after intravenous administration of HAR and HAL both of 1.0mg/kg. The main pharmacokinetic parameters of Cmax, Vd, CL, AUC and MRT, except Ke and t1/2 values, showed significant difference between the two parent drug HAR and HAL, respectively (p<0.05-0.001). Because of the different metabolic rate of HAR and HAL in vivo, the two metabolites, HOL and HAM, exhibited unique pharmacokinetic properties.

Topics: Animals; Chromatography, High Pressure Liquid; Dogs; Female; Harmaline; Harmine; Male; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry

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.

Topics: Animals; Antimalarials; Artemisinins; Chloroquine; Drug Synergism; Harmaline; Harmine; HSP90 Heat-Shock Proteins; Malaria; Mice; Mice, Inbred BALB C; Mutagenesis, Site-Directed; Plasmodium berghei; Plasmodium falciparum; Protein Structure, Quaternary; Protein Structure, Tertiary; Surface Plasmon Resonance

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.

Topics: Animals; Antineoplastic Agents; Binding Sites; Carbolines; Cattle; DNA; Harmaline; Harmine; Intercalating Agents; Nucleic Acid Conformation; Peganum; Spectrophotometry, Ultraviolet; Spectroscopy, Fourier Transform Infrared

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.

Topics: Carbolines; Harmaline; Harmine; Nucleic Acid Conformation; Peganum; Plants, Medicinal; RNA, Fungal

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.

Topics: Chromatography, High Pressure Liquid; Flowers; Fructose; Glucose; Harmaline; Harmine; Peganum; Sucrose

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.

Topics: Adenocarcinoma; Apoptosis; Carcinoma, Adenosquamous; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Caspase 3; Caspase 8; Caspase 9; Cell Line, Tumor; Enzyme Activation; Fas Ligand Protein; fas Receptor; Harmaline; Harmine; Humans; Lung Neoplasms; Neoplasm Proteins

In the present study the antimicrobial potential of various extracts from 12 medicinal plants has been investigated in vitro on multiple antibiotic resistant pathogens and some selected protozoa isolated from poultry. The initial examination was performed on E. coli (n = 10) using disc and agar well diffusion assays. Only two plants, Peganum harmala (seeds) and Eucalyptus globulus (leaves) showed positive responses. The active extracts were also investigated against an additional 19 bacteria and the clonal cultures of three protozoa (Histomonas meleagridis, Tetratrichomonas gallinarum and Blastocystis sp.). Only Peganum harmala was found to inhibit the growth of all bacteria and protozoa at 0.38-1.55 mg/mL and 0.63-1.65 mg/mL, respectively. To investigate the potential role of alkaloids in crude extracts of Peganum harmala, four known beta-carbolin alkaloids were quantified and their antimicrobial activity was compared using a microdilution assay. Harmaline was found to be in the highest concentration followed by harmine and harmalol, whereas harmane could not be detected. The activity of the pure alkaloids was in the order harmane > harmaline > harmalol > or = harmine for all bacteria, while for protozoa, it was different depending on the microorganism. It is concluded that Peganum harmala or its alkaloids could probably be used for the control of antibiotic resistant isolates of bacteria as well as protozoa.

Topics: Animals; Anti-Bacterial Agents; Antiprotozoal Agents; Bacteria; Carbolines; Chickens; Drug Resistance; Eukaryota; Harmaline; Harmine; Microbial Sensitivity Tests; Parasitic Sensitivity Tests; Peganum; Phytotherapy; Plant Extracts; Plants, Medicinal

A simple and sensitive method for separation and determination of harmol, harmalol, harmine and harmaline has been developed and validated. Harmol, harmalol, harmine and harmaline were separated using a Metasil ODS column by isocratic elution with flow rate 1.5 ml/min. The mobile phase composition was Isopropyl alcohol-Acetonitrile-Water-Formic acid (100:100:300:0.3) (v/v/v/v) and pH adjusted 8.6 with triethylamine. Spectrophotometric detection was carried out at 330 nm. The linear range of detection for harmol, harmalol, harmine and harmaline were between 9.375-250, 30.750-246, 31.250-500 and 31.000-248 microg/ml, respectively. The method described was suitable for the determination of harmol, harmalol, harmine and harmaline in the seeds of Peganum harmala L.

Topics: Chromatography, High Pressure Liquid; Harmaline; Harmine; Peganum; Reproducibility of Results; Seeds; Sensitivity and Specificity; Spectrophotometry, Ultraviolet

Several microorganisms showed the ability to transform the harman alkaloids, harmaline (1), harmalol (2) and harman (5). Harmaline (1) and harmalol (2) were converted by Rhodotorula rubra ATCC 20129 into the tryptamines, 2-acetyl-3-(2-acetamidoethyl)-7-methoxyindole (3) and 2-acetyl-3-(2-acetamidoethyl)-7-hydroxyindole (4), respectively. Harman (5) was biotransformed by Cunninghamella echinulata NRRL 3655 into 6-hydroxyharman (6) and harman-2-oxide (7).

Topics: Biotransformation; Cunninghamella; Harmaline; Harmine; Magnetic Resonance Spectroscopy; Molecular Structure; Rhodotorula

Three psychological active principles from the seeds of Peganum harmala L., harmine, harmaline and harmalol, showed vasorelaxant activities in isolated rat thoracic aorta preparations precontracted by phenylephrine or KCl with rank order of relaxation potency of harmine > harmaline > harmalol. The vasorelaxant effects of harmine and harmaline (but not harmalol) were attenuated by endothelium removal or pretreatment with a nitric oxide (NO) synthase Nomega-nitro-L-arginine methyl ester. In cultured rat aortic endothelial cells, harmine and harmaline (but not harmalol) increased NO release, which was dependent on the presence of external Ca2+. In endothelium-denuded preparations, pretreatment of harmine, harmaline or harmalol (3-30 microM) inhibited phenylephrine-induced contractions in a non-competitive manner. Receptor binding assays indicated that all 3 compounds interacted with cardiac alpha1-adrenoceptors with comparable affinities (Ki value around 31 - 36 microM), but only harmine weakly interacted with the cardiac 1,4-dihydropyridine binding site of L-type Ca2+ channels (Ki value of 408 microM). Therefore, the present results suggested that the vasorelaxant effects of harmine and harmaline are attributed to their actions on the endothelial cells to release NO and on the vascular smooth muscles to inhibit the contractions induced by the activation of receptor-linked and voltage-dependent Ca2+ channels. The vasorelaxant effect of harmalol was not endothelium-dependent.

Topics: Animals; Aorta, Thoracic; Calcium Channels, L-Type; Cells, Cultured; Dihydropyridines; Endothelium, Vascular; Harmaline; Harmine; In Vitro Techniques; Muscle Contraction; Muscle, Smooth, Vascular; Myocardium; Rats; Rats, Sprague-Dawley; Receptors, Adrenergic, alpha-1; Vasodilator Agents

Several beta-carboline (9H-pyrido-[3,4-b]-indole) alkaloids were evaluated for in vitro trypanosomicidal activity against Trypanosoma cruzi epimastigotes belonging to two different strains (Tulahuén and LQ) showing different sensitivity to nifurtimox. Important differences were observed in the susceptibility of the parasites to these natural substances, with the relatively nifurtimox-resistant LQ strain showing greater sensitivity to the beta-carbolines. Respiratory chain inhibition appears to be a possible determinant of the trypanosomicidal activity of these compounds.

Topics: Animals; Carbolines; Harmaline; Harmine; Kinetics; Nifurtimox; Oxygen Consumption; Species Specificity; Trypanocidal Agents; Trypanosoma cruzi

Topics: 12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; Arachidonic Acid; Blood Platelets; Harmaline; Harmine; Humans; Hydroxyeicosatetraenoic Acids; In Vitro Techniques; Platelet Aggregation; Platelet Aggregation Inhibitors; Thromboxane A2

beta-Carboline alkaloids are derived as a result of condensation between indoleamine (e.g. tryptamine) and short-chain carboxylic acid (e.g. pyruvic acid) or aldehyde (e.g. acetaldehyde), a reaction that occurs readily at room temperature. These compounds have been found endogenously in human and animal tissues and may be formed as a byproduct of secondary metabolism: their endogenous functions however, are not well understood. Indoles and tryptophan derivatives exhibit antioxidative actions by scavenging free radicals and forming resonance stabilized indolyl radicals. Harmane and related compounds exhibited concentration-dependent inhibition of lipid peroxidation (measured as thiobarbiturate reactive products) in a hepatic microsomal preparation incubated with either enzymatic dependent (Fe3+ ADP/NADPH) or non-enzymatic dependent (Fe3+ ADP/dihydroxyfumarate) oxygen radical producing systems. Alkaloids with hydroxyl substitution and a partially desaturated pyridyl ring were found to have the highest antioxidative potencies. Substitution of a hydroxyl group by a methoxyl group at the 6-position resulted in a decrease of greater than 10-fold in the antioxidative activities. Harmane showed high efficacy in an enzymatic system but low efficacy in a non-enzymatic system. The antioxidative effects of harmane in the former system may be attributed to its ability to inhibit oxidative enzymes in the microsomal system. These results suggest that beta-carbolines may also serve as endogenous antioxidants.

Topics: Animals; Antioxidants; Carbolines; Dose-Response Relationship, Drug; Free Radicals; Harmaline; Harmine; Lipid Peroxidation; Male; Microsomes, Liver; Models, Chemical; Rats; Rats, Inbred Strains; Structure-Activity Relationship; Thiobarbiturates; Tryptamines

Room temperature electronic absorption and fluorescence spectra of harmol, harmalol and 2-hydroxycarbazole have been obtained in concentrated aqueous potassium hydroxide solutions. The appearance of a new fluorescence band for all these compounds in media of H- greater than 16, has been ascribed to the emission of excited dianions formed by deprotonation. Acidity constants have been estimated from the Föster-Weller method.

Topics: Carbazoles; Harmaline; Harmine; Hydrogen-Ion Concentration; Hydroxides; Potassium; Potassium Compounds; Solutions; Spectrometry, Fluorescence; Spectrophotometry, Ultraviolet

The preparation of novel biologically active platinum(II) and palladium(II) complexes of some beta-carboline alkaloids (harmaline, harmalol, harmine, and harmane) is described. These complexes, characterized on the basis of their CHN elemental analysis, infrared, Raman and 1H and 13C nuclear resonance spectral data, were shown to have the empirical formula [M(alkaloid)Cl2], M = Pt, Pd. The antitumor and antiviral activities of some of these complexes have been demonstrated.

Topics: Alkaloids; Antineoplastic Agents; Antiviral Agents; Carbolines; Harmaline; Harmine; Magnetic Resonance Spectroscopy; Molecular Structure; Organometallic Compounds; Organoplatinum Compounds; Palladium; Spectrophotometry, Infrared; Spectrum Analysis, Raman; Tumor Cells, Cultured

The ionization and UV-visible spectral properties of some harmala alkaloids have been investigated spectrophotometrically. Harmaline and harmine were found to have pKa values of 9.55 +/- 0.04 and 7.45 +/- 0.03, respectively. The ionization of harmalol was characterized by two processes which could be spectrophotometrically isolated from one another, allowing pKa values of 8.62 +/- 0.15 and 11.30 +/- 0.23 to be determined. The lower of these was ascribed to the phenolic group and the higher to the enamino site. Support for this assignment lay in the yellow color (lambda max 433 nm) at intermediate pH values, which was typical of a formally neutral quinone-methide structure. For harmol, pKa values of 7.90 and 9.47, reported at 21 degrees by Perrin [N.Z.J. Sci. Technol. 388:688-694 (1957)], were reassigned with the lower value reflecting phenolic ionization, as opposed to the original literature assignment. Partition coefficients at pH 7.4 (n-heptane/water) were determined. The comparative pharmacology of these alkaloids is discussed and related to both their pKa values and the relative stability of the neutral quinone-methide structure.

Topics: Alkaloids; Harmaline; Harmine; Hydrogen-Ion Concentration; Ions; Kinetics; Spectrophotometry, Ultraviolet; Structure-Activity Relationship

Harmaline, harmine, 2-methylharmine and harmolol, members of the harmala family of alkaloids were tested for their action on ouabain-elicited contractions of the guinea-pig ileum. All four alkaloids were found to relax tissue previously contracted by ouabain, but they differed in their potency and reversibility. Harmaline was equipotent with harmine but more easily reversible. 2-Methylharmine and harmolol were found to be less potent and readily reversible. It is evident that the inhibition mediated by harmaline occurs at a site other than that which ouabain binds. Ca-free Tyrode caused relaxation of tissue contracted by ouabain. This effect was partially reversed in the presence of normal Tyrode solution. In presence of high K the response to ouabain was almost abolished. Harmaline inhibited Ca-induced contractions in a non-competitive manner while neither 2-methylharmine nor harmalol had any effect. In the discussion the action of harmala alkaloids is shown to be directly related with CA ion movements.

Topics: Alkaloids; Animals; Calcium; Dose-Response Relationship, Drug; Female; Guinea Pigs; Harmaline; Harmine; Ileum; Muscle Contraction; Muscle, Smooth; Ouabain

The actions of five harmala alkaloids on the sodium dependent high affinity choline uptake activity in rat striatal synaptosomes was investigated. All five compounds were found to be competitive inhibitors of the uptake system. Harmalol (Ki approximately 3.4 microM) and 2-methylharmine (Ki approximately 5.7 microM) were found to be relatively potent inhibitors in a series with an ascending order of inhibitory potency of harmaline less than 2-methylharmaline less than harmine less than 2-methylharmine less than harmalol.

Topics: Alkaloids; Animals; Binding Sites; Choline; Corpus Striatum; Harmaline; Harmine; In Vitro Techniques; Male; Rats; Rats, Inbred Strains; Sodium; Synaptosomes

Each of three harmala alkaloids, harmine, harmaline, and harmalol, decreased heart rate and increased pulse pressure, peak aortic flow, and myocardial contractile force in intact normotensive anesthetized dogs. Harmine reduced systemic arterial blood pressure and total peripheral vascular resistance; harmaline-evoked decreases were frequently followed by a secondary increase; and the effects of harmalol on these two parameters were inconsistent. A direct negative chronotropic effect of harmala alkaloids was suggested by observations of bradycardia in the isolated perfused rat heart and in the intact dog; neither vagotomy nor atropinization affected harmala alkaloid-induced bradycardia in the dog. Reduction in femoral vascular resistance by the alkaloids was not apparently due to activation of cholinergic, beta-adrenergic, or histamine (H1) receptors.

Topics: Alkaloids; Animals; Atropine; Blood Pressure; Dogs; Female; Harmaline; Harmine; Heart Rate; Hemodynamics; Hindlimb; In Vitro Techniques; Male; Myocardial Contraction; Rats; Vagotomy; Vascular Resistance