Β-carboline Harmine Research Articles

Hair analysis to document ayahuasca consumption by LC-MS/MS

Ayahuasca is a hallucinogenic drink consumed during shamanic rituals for spiritual and curative purposes. In recent years, the use of this beverage was no longer restricted to the Amazon rainforest but has spread all over the world and notably in Europe. The internationalisation of Ayahuasca raises risks associated with its consumption during rituals performed by individuals lacking in knowledge of the psychotropic ingredients and who could abuse the situation of an emotional and a physical vulnerability. To document exposure to ayahuasca, hair analysis could be very helpful, with application in both clinical and historical contexts to evidence the consumption of this substance. This can also be done on archaeological specimens. In this study, the hair of a subject suspected of ayahuasca consumption were analysed using an original LC-MS/MS method. The three β-carbolines harmine (2.5–1,300 pg/mg), harmaline (30–89,700 pg/mg) and tetrahydroharmine (2.2–2.5 pg/mg) and the tryptamines 5-OH-DMT (2–62 pg/mg) and 5-MeO-DMT (< LOQ) tested positive. Dimethyltryptamine tested negative.For the first time, values of all β-carbolines and 5-OH-DMT in hair are displayed in the literature. The absence of dimethyltryptamine in the hair proves that the analytical strategy to be used to evidence ayahuasca consumers should not be directed at the single research of DMT.Since there is not a single ayahuasca recipe, it will be necessary to test not only DMT but also the other ayahuasca alkaloids such as β-carbolines (harmine, harmaline and THH) and other tryptamines (5-OH-DMT and 5-MeO-DMT) to fully document exposure.

The β-Carboline Harmine Has a Protective Immunomodulatory Role in Nonhealing Cutaneous Leishmaniasis

Cutaneous leishmaniasis affects 1 million people worldwide annually. Although conventional treatments primarily target the parasite, there is growing interest in host immune modulation. In this study, we investigated the impact of synthetic β-carboline harmine (ACB1801), previously shown to be immunoregulatory in cancer, on the pathology caused by a drug-resistant Leishmania major strain causing persistent cutaneous lesions. Exposure to ACB1801 invitro had a modest impact on parasite burden within host macrophages. Moreover, it significantly increased major histocompatibility complex II and costimulatory molecule expression on infected dendritic cells, suggesting an enhanced immune response. Invivo, ACB1801 monotherapy led to a substantial reduction in lesion development and parasite burden in infected C57BL/6 mice, comparable with efficacy of amphotericin B. Transcriptomics analysis further supported ACB1801 immunomodulatory effects, revealing an enrichment of TNF-α, IFN-γ, and major histocompatibility complex II antigen presentation signatures in the draining lymph nodes of treated mice. Flow cytometry analysis confirmed an increased frequency (1.5×) of protective CD4+IFN-γ+TNF-α+ T cells and a decreased frequency (2×) in suppressive IL-10+FoxP3- T cells at the site of infection and in draining lymph nodes. In addition, ACB1801 downregulated the aryl hydrocarbon receptor signaling, known to enhance immunosuppressive cytokines. Thus, these results suggest a potential use for ACB1801 alone or in combination therapy for cutaneous leishmaniasis.

Harmine impairs memory performance of treated rats and nontreated cagemates.

The interest in psychedelic substances as potential treatments for psychiatric disorders is increasing. The β-carboline harmine, an Ayahuasca component, presents hallucinogenic and antidepressant effects. Although Ayuahuasca-and consequently harmine-is usually consumed in rituals, the role of social contexts in the behavioral effects of harmine has not been investigated yet. In this sense, affective states may modulate cohabitants' behavior, including learning/memory. This work investigates the effects of harmine on the learning/memory performance of rats evaluated on the contextual and tone fear conditioning (CFC and TFC) and on the plus-maze discriminative avoidance (PMDAT) tasks. The possible influence of a harmine-treated cohabitant was assessed by evaluating rats housed in homogeneous cages-where all the animals were acutely administered with the same treatment (vehicle, 5, 10, or 15 mg/kg harmine), and in heterogeneous cages-where each animal received a different drug treatment. The main results are: (a) harmine impaired CFC (10 mg/kg) and PMDAT discrimination (all doses); and (b) harmine caused a memory deficit in CFC, TFC, and PMDAT of untreated rats kept in heterogeneous cages. Our results show that harmine induces a memory deficit in tasks with emotional contexts. Further, the cohabitation with animals treated with this drug also seems to impair memory performance of untreated animals. (PsycInfo Database Record (c) 2022 APA, all rights reserved).

The β-carboline Harmine improves the therapeutic benefit of anti-PD1 in melanoma by increasing the MHC-I-dependent antigen presentation.

Harmine is a dual-specificity tyrosine-regulated kinase 1A (DYRK1A) inhibitor that displays a number of biological and pharmacological properties. Also referred to as ACB1801 molecule, we have previously reported that harmine increases the presentation of major histocompatibility complex (MHC)-I-dependent antigen on melanoma cells. Here, we show that ACB1801 upregulates the mRNA expression of several proteins of the MHC-I such as Transporter Associated with antigen Processing TAP1 and 2, Tapasin and Lmp2 (hereafter referred to as MHC-I signature) in melanoma cells. Treatment of mice bearing melanoma B16-F10 with ACB1801 inhibits the growth and weight of tumors and induces a profound modification of the tumor immune landscape. Strikingly, combining ACB1801 with anti-PD1 significantly improves its therapeutic benefit in B16-F10 melanoma-bearing mice. These results suggest that, by increasing the MHC-I, ACB1801 can be combined with anti-PD1/PD-L1 therapy to improve the survival benefit in cancer patients displaying a defect in MHC-I expression. This is further supported by data showing that i) high expression levels of TAP1, Tapasin and Lmp2 was observed in melanoma patients that respond to anti-PD1; ii) the survival is significantly improved in melanoma patients who express high MHC-I signature relative to those expressing low MHC-I signature; and iii) high expression of MHC-I signature in melanoma patients was correlated with increased expression of CD8 and NK cell markers and overexpression of proinflammatory chemokines involved in the recruitment of CD8+ T cells.

Kinetic profile of N,N-dimethyltryptamine and β-carbolines in saliva and serum after oral administration of ayahuasca in a religious context.

Ayahuasca is a beverage obtained from Banisteriopsis caapi plus Psychotria viridis. B. caapi contains the β-carbolines harmine, harmaline, and tetrahydroharmine that are monoamine oxidase inhibitors and P. viridis contains N,N-dimethyltryptamine (DMT) that is responsible for the visionary effects of the beverage. Ayahuasca use is becoming a global phenomenon, and the recreational use of DMT and similar alkaloids has also increased in recent years; such uncontrolled use can lead to severe intoxications. In this investigation, liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to study the kinetics of alkaloids over a 24 h period in saliva and serum of 14 volunteers who consumed ayahuasca twice a month in a religious context. We compared the area under the curve (AUC), maximum concentration (Cmax ), time to reach Cmax (Tmax ), mean residence time (MRT), and half-life (t1/2 ), as well as the serum/saliva ratios of these parameters. DMT and β-carboline concentrations (Cmax ) and AUC were higher in saliva than in serum and the MRT was 1.5-3.0 times higher in serum. A generalized estimation equations (GEEs) model suggested that serum concentrations could be predicted by saliva concentrations, despite large individual variability in the saliva and serum alkaloid concentrations. The possibility of using saliva as a biological matrix to detect DMT, β-carbolines, and their derivatives is very interesting because it allows fast noninvasive sample collection and could be useful for detecting similar alkaloids used recreationally that have considerable potential for intoxication.

The β-Carboline Harmine Induces Actin Dynamic Remodeling and Abrogates the Malignant Phenotype in Tumorigenic Cells.

Numerous studies have shown that alteration of actin remodeling plays a pivotal role in the regulation of morphologic and phenotypic changes leading to malignancy. In the present study, we searched for drugs that can regulate actin polymerization and reverse the malignant phenotype in cancer cells. We developed a cell-free high-throughput screening assay for the identification of compounds that induce the actin polymerization in vitro, by fluorescence anisotropy. Then, the potential of the hit compound to restore the actin cytoskeleton and reverse the malignant phenotype was checked in EWS-Fli1-transformed fibroblasts and in B16-F10 melanoma cells. A β-carboline extracted from Peganum harmala (i.e., harmine) is identified as a stimulator of actin polymerization through a mechanism independent of actin binding and requiring intracellular factors involved in a process that regulates actin kinetics. Treatment of malignant cells with non-cytotoxic concentrations of harmine induces the recovery of a non-malignant cell morphology accompanied by reorganization of the actin cytoskeleton, rescued cell–cell adhesion, inhibition of cell motility and loss of anchorage-independent growth. In conclusion, harmine induces the reversion of the malignant phenotype by a process involving the modulation of actin dynamics and is a potential anti-tumor agent acting principally through a non-cytotoxic process.

Harmicines − harmine and cinnamic acid hybrids as novel antiplasmodial hits

Harmicines constitute novel hybrid compounds that combine two agents with reported antiplasmodial properties, namely β-carboline harmine and a cinnamic acid derivative (CAD). Cu(I) catalyzed azide-alkyne cycloaddition was employed for the preparation of three classes of hybrid molecules: N-harmicines 6a-i, O-harmicines 7a-i and N,O-bis-harmicines 8a-g,i. In vitro antiplasmodial activities of harmicines against the erythrocytic stage of Plasmodium falciparum (chloroquine-sensitive Pf3D7 and chloroquine-resistant PfDd2 strains) and hepatic stage of P. berghei, as well as cytotoxicity against human liver hepatocellular carcinoma cell line (HepG2), were evaluated. Remarkably, most of the compounds exerted significant activities against both stages of the Plasmodium life cycle. The conjugation of various CADs to harmine resulted in the increased antiplasmodial activity relative to harmine. In general, O-harmicines 7 exhibited the highest activity against the erythrocytic stage of both P. falciparum strains, whereas N,O-bis harmicines 8 showed the most pronounced activity against P. berghei hepatic stages. For the latter compound, molecular dynamics simulations confirmed binding within the ATP binding site of PfHsp90, while the weaker binders, namely 6b and harmine, were found to be positioned away from this structural element. In addition, decomposition of the computed binding free energies into contributions from individual residues suggested guidelines for further derivatization of harmine towards more efficient compounds. Cytotoxicity screening revealed N-harmicines 6 as the least, and O-harmicines 7 as the most toxic compounds. Harmicines 6g, 8b and 6d exerted the most selective action towards Plasmodium over human cells, respectively. These results establish harmicines as hits for future optimisation and development of novel antiplasmodial agents.

Identification of Plant-derived Alkaloids with Therapeutic Potential for Myotonic Dystrophy Type I

Myotonic dystrophy type I (DM1) is a disabling neuromuscular disease with no causal treatment available. This disease is caused by expanded CTG trinucleotide repeats in the 3' UTR of the dystrophia myotonica protein kinase gene. On the RNA level, expanded (CUG)n repeats form hairpin structures that sequester splicing factors such as muscleblind-like 1 (MBNL1). Lack of available MBNL1 leads to misregulated alternative splicing of many target pre-mRNAs, leading to the multisystemic symptoms in DM1. Many studies aiming to identify small molecules that target the (CUG)n-MBNL1 complex focused on synthetic molecules. In an effort to identify new small molecules that liberate sequestered MBNL1 from (CUG)n RNA, we focused specifically on small molecules of natural origin. Natural products remain an important source for drugs and play a significant role in providing novel leads and pharmacophores for medicinal chemistry. In a new DM1 mechanism-based biochemical assay, we screened a collection of isolated natural compounds and a library of over 2100 extracts from plants and fungal strains. HPLC-based activity profiling in combination with spectroscopic methods were used to identify the active principles in the extracts. The bioactivity of the identified compounds was investigated in a human cell model and in a mouse model of DM1. We identified several alkaloids, including the β-carboline harmine and the isoquinoline berberine, that ameliorated certain aspects of the DM1 pathology in these models. Alkaloids as a compound class may have potential for drug discovery in other RNA-mediated diseases.

β-Carboline harmine reverses the effects induced by stress on behaviour and citrate synthase activity in the rat prefrontal cortex.

The present study was aimed at evaluating the effects of the administration of β-carboline harmine on behaviour and citrate synthase activity in the brain of rats exposed to chronic mild stress (CMS) procedure. To this aim, after 40 days of exposure to CMS procedure, rats were treated with harmine (15 mg/kg/day) for 7 days, then memory, anhedonia and citrate synthase activity were assessed. Result Our findings demonstrated that stressed rats treated with saline increased the sucrose intake, and the stressed rats treated with harmine reversed this effect. Neither stress nor harmine treatment altered memory performance in rats. In addition, chronic stressful situations induced increase in citrate synthase activity in the prefrontal cortex, but not in the hippocampus and striatum. Treatment with harmine reversed the increase in citrate synthase activity in the prefrontal cortex. These findings support the hypothesis that harmine could be involved in controlling the energy metabolism.

Chronic administration of harmine elicits antidepressant-like effects and increases BDNF levels in rat hippocampus

A growing body of evidence has pointed to the β-carboline harmine as a potential therapeutic target for the treatment of major depression. The present study was aimed to evaluate behavioural and molecular effects of the chronic treatment with harmine and imipramine in rats. To this aim, rats were treated for 14 days once a day with harmine (5, 10 and 15 mg/kg) and imipramine (10, 20 and 30 mg/kg) and then subjected to the forced swimming and open-field tests. Harmine and imipramine, at all doses tested, reduced immobility time of rats compared with the saline group. Imipramine increased the swimming time at 20 and 30 mg/kg and harmine increased swimming time at all doses. The climbing time increased in rats treated with imipramine (10 and 30 mg/kg) and harmine (5 and 10 mg/kg), without affecting spontaneous locomotor activity. Brain-derived neurotrophic factor (BDNF) hippocampal levels were assessed in imipramine and harmine-treated rats by ELISA sandwich assay. Interestingly, chronic administration of harmine at the higher doses (10 and 15 mg/kg), but not imipramine, increased BDNF protein levels in rat hippocampus. Finally, these findings further support the hypothesis that harmine could bring about behavior and molecular effects, similar to antidepressants drugs.

Harmine and imipramine promote antioxidant activities in prefrontal cortex and hippocampus.

A growing body of evidence has suggested that reactive oxygen species (ROS) may play an important role in the physiopathology of depression. Evidence has pointed to the β-carboline harmine as a potential therapeutic target for the treatment of depression. The present study we evaluated the effects of acute and chronic administration of harmine (5, 10 and 15 mg/kg) and imipramine (10, 20 and 30 mg/kg) or saline in lipid and protein oxidation levels and superoxide dismutase (SOD) and catalase (CAT) activities in rat prefrontal cortex and hippocampus. Acute and chronic treatments with imipramine and harmine reduced lipid and protein oxidation, compared to control group in prefrontal cortex and hippocampus. The SOD and CAT activities increased with acute and chronic treatments with imipramine and harmine, compared to control group in prefrontal cortex and hippocampus. In conclusion, our results indicate positive effects of imipramine antidepressant and β-carboline harmine of oxidative stress parameters, increasing SOD and CAT activities and decreasing lipid and protein oxidation.

Effects of β-carboline harmine on behavioral and physiological parameters observed in the chronic mild stress model: Further evidence of antidepressant properties

The chronic mild stress (CMS) model has been used as an animal model of depression which induces anhedonic behavior in rodents. The present study was aimed to evaluate the behavioral and physiological effects of administration of β-carboline harmine in rats exposed to CMS procedure. To this aim, after 40 days of exposure to CMS procedure, rats were treated with harmine (15 mg/kg/day) for 7 days. In this study, sweet food consumption, adrenal gland weight, adrenocorticotrophin hormone (ACTH) levels, and hippocampal brain-derived-neurotrophic factor (BDNF) protein levels were assessed. Our findings demonstrated that chronic stressful situations induced anhedonia, hypertrophy of adrenal gland weight, increase ACTH circulating levels in rats and increase BDNF protein levels. Interestingly, treatment with harmine reversed anhedonia, the increase of adrenal gland weight, normalized ACTH circulating levels and BDNF protein levels. Finally, these findings further support the hypothesis that harmine could be a new pharmacological tool for the treatment of depression.

Behavioral profile of constituents in ayahuasca, an Amazonian psychoactive plant mixture

Ayahuasca is a psychoactive plant mixture typically composed of the β-carboline-rich Banisteriopsis caapi vine and the hallucinogenic plant Psychotria viridis. Ayahuasca has long been used by aboriginal populations for its putative spiritual and medicinal benefits. Although the presumed primary chemical constituents of ayahuasca have been identified, little is known about the basic in vivo pharmacology of the extract. Two principal constituents of ayahuasca, the β-carboline harmine and N, N-dimethyltryptamine (DMT) were selected for detailed study in mice using the Functional Observational Battery (FOB). The B. caapi extract was then examined alone and in combination with DMT. Harmine and the B. caapi extract produced similar effects in the FOB, particularly in the open field. Clonic and tonic motor movements were augmented by DMT administration. Harmine and B. caapi decreased acoustic startle amplitude without significantly affecting prepulse inhibition. DMT appeared to attenuate startle-decreasing effects of harmine and B. caapi, although these effects fell just short of significance. These results suggest that the behavioral effects of B. caapi in mice may be attributed in large part to its principal alkaloid species, harmine, and related β-carbolines in the extract. Hence, the presence of the banisteriopsis vine in the admixture may directly contribute to the unique subjective effects of ayahuasca.

Spectrofluorimetric Study of the Effects of Cyclodextrins on the Acid–Base Equilibria of Harmine and Harmane

β-Carboline alkaloids are important compounds because they exhibit a variety of pharmacological actions. Their acid–base behaviour can be studied by spectrofluorimetry since these molecules present a remarkable native luminescence. Acid–base equilibria depend on the environment of the molecules and inclusion into cyclodextrin (CD) cavities shifts the acid–base equilibria and alters the apparent pK a values. The influence of CDs on the acid–base equilibria of the model β-carbolines harmine and harmane is described. β-CD and γ-CD and the modified β-CDs hydroxypropyl-β-CD (HPβ-CD), 2,6-di-O-methyl-β-CD (DMβ-CD) and 2,3,6-tri-O-methyl-β-CD (TMβ-CD) were used to form the corresponding complexes with harmine and harmane in the pH range 7.8–8.0. In these buffered solutions the complexes with the different CDs exhibit an emission band with resolved peaks at 360 and 380 nm corresponding to the neutral form of harmane and with a remarkable enhancement in the emission intensity compared with aqueous solution. In the case of the complexes with β-CD and γ-CD, both the cationic and the neutral emission bands appear. However, for γ-CD the cationic band is more intense than the neutral band, the inverse being true for β-CD. In homogeneous aqueous solution at this pH value the cationic band is the only one observed and therefore the presence of the neutral band indicates the formation of inclusion complexes. In the harmane–HPβ-CD complexes, the emission bands ascribed to the anionic form are observed after addition of NaOH. This emission is only observed in homogeneous aqueous solution in strongly alkaline media outside the normal pH range.

Harmine reverses the inhibition of lordosis by the 5-HT 2 antagonists pirenperone and ketanserin in the female rat

The β-carboline harmine was found to facilitate lordosis behavior in ovariectomized rats primed with estradiol benzoate. Moreover, harmine reversed the inhibition of lordosis by the serotonin type 2 (5-HT 2) antagonists pirenperone and ketanserin in rats primed with estradiol benzoate and progesterone. These results suggest that harmine facilities lordosis by enhancing activity at 5-HT 2 receptors.

Differential effects of phenobarbitone and 3-methylcholanthrene induction on the hepatic microsomal metabolism of the β-carbolines harmine and harmol

The metabolism of the β-carbolines harmine and harmol by C57/BL10 mouse liver microsomes is reported. Marked changes in apparent Michaelis-Menten kinetics and metabolite profiles were induced differentially by phenobarbitone (PB) or 3-methylcholanthrene (MC). With control or PB-induced microsomes harmine was metabolised by both high- and low-affinity reactions (app. K m = 1 and 29 μM for controls; app. K m = 1.9 and 21 μM for PB-induced). Only the high-affinity reaction occurred following MC pretreatment, induced 31-fold to V max = 22 nmoles·min −1·mg protein −1 (app. K m = 0.4 μM). With control or PB-induced microsomes harmine was metabolised almost exclusively by O-demethylation to harmol (3-fold induction to V max = 9.8 nmoles·min −1·mg protein −1, app. K m = 73 and 14 μM respectively), which was probably the low-affinity reaction of harmine metabolism. With MC-induced microsomes harmine was metabolised mainly to two unidentified yellow products, with harmol as a minor metabolite formed by a high-affinity reaction (app. K m = 0.6 μM). Control, PB- and MC-induced microsomes each metabolised harmol by a high-affinity reaction (app. K m = 0.6–1.9 μM) to unidentified metabolites. Harmol metabolism was not induced by PB, but was induced by MC (22-fold to Vmax = 11 nmoles·min −1·g protein −1): this partly explains the relative lack of net harmol production after MC induction. The unidentified MC-induced yellow metabolites of harmine were not formed via harmol. Harmine and harmol metabolism showed the characteristics of cytochrome P-450 catalysed reactions. Harmine gave Type II cytochrome P-450 binding spectra with control and PB-induced microsomes, but a high-affinity Type I spectrum ( K s = 6.5 μM) with MC-induced microsomes. Harmol gave modified Type II spectra in all cases, with again higher-affinity binding to MC-induced microsomes ( K s = 62 μM) than to control ( K s ⪢ 500 μM) or PB-induced microsomes ( K s = 500 μM).