ergoline and elymoclavine

Nowadays psychoactive plants marketed as "legal highs" or "herbal highs" increase in popularity. One popular "legal high" are the seeds of the Hawaiian baby woodrose Argyreia nervosa (Synonym: Argyreia speciosa, Convolvolus speciosus). At present there exists no study on A. nervosa seeds or products, which are used by consumers. The quality of commercial available A. nervosa seeds or products is completely unknown. In the present study, a commercial available seed collection (five seeds labeled "flash of inspiration", FOI) was analyzed for ergot alkaloids together with an A. nervosa product (two preparations in capsule form, "druids fantasy", DF). For this purpose high performance liquid chromatography high resolution tandem mass spectrometry (HPLC-HRMS/MS) technique was employed. Besides the major ingredients such as lysergic acid amide (LSA) and ergometrine the well known A. nervosa compounds lysergol/elymoclavine/setoclavine, chanoclavine and the respective stereoisomers were detected in DF, while only LSA and ergometrine could be found in FOI. In addition, in DF lysergic acid was found, which has not been reported yet as ingredient of A. nervosa. In both products, DF as well as in FOI, LSA/LSA-isomers were dominant with 83-84% followed by ergometrine/ergometrinine with 10-17%. Therefore, LSA, followed by ergometrine/ergometrinine, could be confirmed to be the main ergot alkaloids present in A. nervosa seeds/products whereas the other ergot alkaloids seemed to be of minor importance (less than 6.1% in DF). The total ergot alkaloid amounts varied considerably between DF and FOI by a factor of 8.6 as well as the LSA concentration ranging from 3 μg (lowest amount in one FOI seed) to approximately 34 μg (highest amount in one DF capsule). Among the FOI seeds, the LSA concentration varied from approximately 3-15 μg per seed. Thus, the quality/potency of seeds/preparations depends on the amount of ergot alkaloids and the intensity of an expected trip is totally unpredictable.

Topics: Alkaloids; Chromatography, Liquid; Convolvulus; Ergolines; Ergonovine; Humans; Indoles; Lysergic Acid Diethylamide; Molecular Structure; Psychotropic Drugs; Seeds; Tandem Mass Spectrometry

The grass parasites Claviceps purpurea and Claviceps fusiformis produce ergot alkaloids (EA) in planta and in submerged culture. Whereas EA synthesis (EAS) in C. purpurea proceeds via clavine intermediates to lysergic acid and the complex ergopeptines, C. fusiformis produces only agroclavine and elymoclavine. In C. purpurea the EAS gene (EAS) cluster includes dmaW (encoding the first pathway step), cloA (elymoclavine oxidation to lysergic acid), and the lpsA/lpsB genes (ergopeptine formation). We analyzed the corresponding C. fusiformis EAS cluster to investigate the evolutionary basis for chemotypic differences between the Claviceps species. Other than three peptide synthetase genes (lpsC and the tandem paralogues lpsA1 and lpsA2), homologues of all C. purpurea EAS genes were identified in C. fusiformis, including homologues of lpsB and cloA, which in C. purpurea encode enzymes for steps after clavine synthesis. Rearrangement of the cluster was evident around lpsB, which is truncated in C. fusiformis. This and several frameshift mutations render CflpsB a pseudogene (CflpsB(Psi)). No obvious inactivating mutation was identified in CfcloA. All C. fusiformis EAS genes, including CflpsB(Psi) and CfcloA, were expressed in culture. Cross-complementation analyses demonstrated that CfcloA and CflpsB(Psi) were expressed in C. purpurea but did not encode functional enzymes. In contrast, CpcloA catalyzed lysergic acid biosynthesis in C. fusiformis, indicating that C. fusiformis terminates its EAS pathway at elymoclavine because the cloA gene product is inactive. We propose that the C. fusiformis EAS cluster evolved from a more complete cluster by loss of some lps genes and by rearrangements and mutations inactivating lpsB and cloA.

Topics: Biosynthetic Pathways; Chromatography, Liquid; Chromatography, Thin Layer; Claviceps; Ergolines; Ergot Alkaloids; Evolution, Molecular; Gene Order; Genes, Fungal; Mass Spectrometry; Models, Biological; Molecular Structure; Multigene Family; Mutation; Species Specificity

Three series of cycloalkanecarboxylic esters derived from the naturally occurring clavine alkaloids lysergol, dihydrolysergol-I, and elymoclavine were synthesized to study their interaction with 5-HT2A receptors and alpha1-adrenoceptors in rat tail artery and aorta, respectively. Especially cycloalkanecarboxylic esters derived from lysergol showed complex behavior as partial agonists and antagonists of the contractile effect of 5-HT. Within this group, partial 5-HT2A receptor agonist activity was most potent for cyclopropanecarboxylic ester 6a (pKP = 7.67, alpha = 0.21) and decreased as the volume requirement of the alicyclic ring increased. This tendency was echoed in experiments where the compounds were used as antagonists of the contractile effect of 5-HT. From the structure-activity study, the N-1-isopropyl homologue of 6a, compound 6b, emerged as the ligand with the highest affinity for rat 5-HT2A receptors (pA2 = 8.74). For cycloalkanecarboxylic esters derived from dihydrolysergol-I and elymoclavine, no clear structure-affinity relationship could be deduced, although those compounds that had smaller cycloalkyl rings in the acyl portion and an isopropyl substituent at N-1 showed the highest 5-HT2A receptor affinity. On the other hand, cycloalkanecarboxylic esters derived from lysergol, dihydrolysergol-I, and elymoclavine displayed low or marginal affinity at alpha1-adrenoceptors. A further aim of the study was to examine to what extent the complete removal of the acyl portion of the esters would affect 5-HT2A receptor affinity. The parent alcohols of the three series of N-1-isopropyl homologues, 1-isopropyllysergol (1b), 1-isopropyldihydrolysergol-I (2b), and 1-isopropylelymoclavine (3b), displayed higher affinity for 5-HT2A receptors (pA2 = 9.15, 8.50, 9.14) than the corresponding esters. Compounds 1b-3b had no contractile effects by themselves and displayed low affinity at guinea-pig 5-HT1B receptors and rat alpha1-adrenoceptors. The high affinity for rat 5-HT2A receptors was retained when clavines even more simple in structure than 1b-3b, compounds 4b and 5b, were examined as 5-HT2A receptor antagonists. The nanomolar antagonist activity of simple clavines (1b-5b) in the rat suggests that the indolo[4,3-fg]quinoline system of the ergolines is the molecular fragment that is responsible for 5-HT2A receptor affinity, and not the substituent at position C-8.

Topics: Animals; Aorta, Thoracic; Arteries; Ergolines; Female; Guinea Pigs; Iliac Artery; In Vitro Techniques; Lysergic Acid; Male; Muscle Contraction; Muscle, Smooth, Vascular; Rats; Rats, Wistar; Receptor, Serotonin, 5-HT1B; Receptor, Serotonin, 5-HT2A; Receptors, Adrenergic, alpha-1; Receptors, Serotonin; Serotonin Antagonists; Serotonin Receptor Agonists; Tail

New glycosides derived from ergot alkaloids elymoclavine and DH-lysergol were synthesized by chemoenzymatic methods. beta-Glucosides were obtained either by chemical method or by transglycosylation (glycosidase from Aspergillus oryzae), lactosides were prepared by further extension of carbohydrate chain using beta-1,4-galactosyltransferase (bovine milk) and alpha-5-N-acetylneuraminyl-(2-->6)-beta-D-galactopyranosyl-(l-->4)-2- acetamido-2-deoxy-beta-D-glucopyranosyl-(1-->O)-elymoclavine was prepared using alpha-2,6-sialyltransferase (rat liver). Immunomodulatory activity of elymoclavine and 9,10-dihydrolysergol and their glycosylated derivatives on natural killer (NK) cell-mediated cytotoxicity of human resting and activated human peripheral blood mononuclear cells (PBMC) was investigated. Addition of ergot alkaloid glycosides to the mixtures of effector and target cells potentiated the PBMC cytotoxicity against both NK-sensitive and -resistant target cells. The glycoconjugates of elymoclavine enhanced cytotoxicity of PBMC against NK-resistant target cells. The glycoconjugates of DH-lysergol potentiated NK cytotoxicity of PBMC against NK-sensitive target cells.

Topics: Adjuvants, Immunologic; Animals; Carbohydrate Sequence; Cells, Cultured; Cytotoxicity, Immunologic; Ergolines; Ergot Alkaloids; Glycosides; Humans; Killer Cells, Natural; Magnetic Resonance Spectroscopy; Molecular Sequence Data; Rats; Spectrometry, Mass, Fast Atom Bombardment; Tumor Cells, Cultured

A series of nine naturally occurring and/or semi-synthetic elymoclavine mono- and oligoglycosides containing various sugar units was investigated by fast atom bombardment, high-resolution measurements and linked scans. The fragmentations of the [M+H]+ and [M+Na]+ ions are described and their different contributions to the structure elucidation are discussed.

Topics: Carbohydrate Sequence; Ergolines; Molecular Sequence Data; Spectrometry, Mass, Fast Atom Bombardment

The interaction between the octapeptide angiotensin II (AT II) and the DA-ergic agents (agonists)--the ergotic alkaloid elymoclavine and bromocryptine--during exploratory behaviour was studied in experiments on male albino rats. The changes in the horizontal and vertical activity of the exploratory behaviour and the hole-board activity were investigated using an Opto-Varimex apparatus. AT II, elymoclavine and bromocryptine were applied alone. The frequency of rearing and ambulation was increased with all substances applied (the effect being most pronounced on the 10th min), while the hole-board activity decreased. Elymoclavine potentiates the effect of AT II during exploratory behaviour. The effects of the drugs tested on the exploratory behaviour most probably result from the interaction between AT II receptors, dopamine receptors and through GABA-ergic neurotransmission, in the respective brain zones responsible for behaviour.

Topics: Angiotensin II; Animals; Behavior, Animal; Bromocriptine; Ergolines; Exploratory Behavior; Male; Motor Activity; Rats; Rats, Inbred Strains

Using the step-down method for passive avoidance with punishment electroshock reinforcement, the experimental rats were divided according to their individual differences in the capacity for learning and retention into two groups: good learners (GL) and poor learners (PL). Stereotypy and catalepsy methods were used in order to clarify the role of the central dopaminergic (DA-ergic) transmitter mechanisms for the individual differences. Stereotypic motor behaviour was induced by applying the following DA-ergic agonists: apomorphine (2 mg/kg. i.p.), amphetamine (6 mg/kg, s.c.) and elymoklavine (2 mg/kg, i.p.). The stereotypic behaviour was recorded on a 5-point scale. A set of catalepsy tests was used to determine the cataleptic effect of the DA-ergic antagonist haloperidol (1 mg/kg, i.p.). Our experimental results showed that the intensity and duration of the stereotypy caused by the three agents were significantly increased in GL compared with PL. Catalepsy in the PL group appeared later than in the GL group, and its total duration was shorter. The data obtained suggest that in the GL group there is a marked tendency towards a higher level of sensitivity of the brain dopamine receptors, compared with the PL group.

Topics: Animals; Apomorphine; Catalepsy; Dopamine; Ergolines; Haloperidol; Learning; Male; Memory; Rats; Rats, Inbred Strains; Reinforcement, Psychology; Stereotyped Behavior; Synaptic Transmission

Topics: Animals; Dose-Response Relationship, Drug; Ergolines; Haloperidol; Male; Rats; Rats, Inbred Strains; Receptors, Dopamine; Stereotyped Behavior

Regulation of the production of clavine alkaloids, especially elymoclavine, by sucrose, maltose, and mixtures of these saccharides was studied in submerged cultures of strains Claviceps purpurea 129/35 and Claviceps sp. SD-58. The data were statistically processed on an EC 1040 computer. Fermentation medium containing sucrose (80 g/l) in combination with glucose (20 g/l) was the best for elymoclavine formation. Retarded release of glucose from maltose increased the formation of elymoclavine and suppressed the synthesis of undesirable extracellular glucans. Carbon source can affect both the total amount of produced alkaloids and the relative proportion of individual clavines in the alkaloid mixture.

Topics: Carbohydrate Metabolism; Claviceps; Ergolines; Glucose; Maltose; Sucrose

The effect of the clavine alkaloid elymoclavine, isolated from Claviceps sp cp II, on the level and turnover of biogenic monoamines in several rat brain structures was studied. Elymoclavine administered intraperitoneally (i.p.) at a dose of 5 mg/kg significantly increased the dopamine (DA) level in the striatum and hypothalamus and enhanced the DA turnover in the striatum. A significant increase was also found in the level and turnover of noradrenaline (NA) in the hypothalamus. Elymoclavine exerted an opposite effect on the level and turnover of serotonin (5-HT) in these brain structures: the 5-HT level significantly declined and the 5-HT turnover slightly decreased in the striatum and hypothalamus. In the cerebral cortex elymoclavine significantly increased the 5-HT level. Although the stimulant effect on DA receptors appears to be a dominant element in the mechanism of action of elymoclavine, it seems that this ergot alkaloid is also characterized by a plurireceptor action. The present results suggest a certain role for the elymoclavine action on the level and turnover of brain biogenic monoamines in the mechanism of its different pharmacological effects.

Topics: Animals; Biogenic Amines; Brain; Cerebral Cortex; Corpus Striatum; Dopamine; Ergolines; Hydroxyindoleacetic Acid; Hypothalamus; Injections, Intraperitoneal; Male; Norepinephrine; Rats; Rats, Inbred Strains; Serotonin

The effect of angiotensin II (ATII) and of its interactions with dopaminergic drugs injected post-trial on retention in active avoidance tasks in shuttle-box-trained rats were studied. ATII at doses of 0.10 and 0.50 micrograms administered intracerebroventricularly (i.c.v.) immediately after training improved retention. The dopaminergic receptor agonist apomorphine at a dose of 0.10 mg/kg injected intraperitoneally (i.p.) facilitated retention whereas elymoclavine (a dopaminergic agonist) at a dose of 2.5 mg/kg i.p. had no effect. ATII at a dose of 0.10 micrograms i.c.v. administered after apomorphine 0.10 mg/kg or elymoclavine 2.5 mg/kg exerted a stronger retention-facilitating effect. The dopaminergic receptor antagonist haloperidol at a dose of 1 mg/kg i.p. markedly impaired retention. ATII at a dose of 0.50 micrograms administered after haloperidol (1 mg/kg) did not exercise its retention-facilitating effect. It is concluded that the retention facilitating effects of ATII are realized through interactions with brain dopaminergic transmission.

Topics: Angiotensin II; Animals; Apomorphine; Avoidance Learning; Dopamine; Drug Interactions; Ergolines; Haloperidol; Male; Memory; Rats; Rats, Inbred Strains; Retention, Psychology

In submerged Claviceps cultures the activity of hydroxymethylglutaryl-CoA reductase preceded the increase of alkaloid production and of sterol content. During the first alkaloid phase, cell mevalonate was involved in the biosynthesis of both alkaloids and steroids. In the second production phase, it was predominantly used for alkaloid synthesis. Hydroxymethylglutaryl-CoA reductase appears to be a suitable target for physiological manipulation to increase clavine alkaloid yields.

Topics: Claviceps; Ergolines; Ergot Alkaloids; Hydroxymethylglutaryl CoA Reductases; Sterols

In training for passive avoidance using a device of the step-down type, the nootropic agents piracetam (60 mg/kg orally) and centrophenoxine (100 mg/kg, i. p.) do not facilitate learning, while the ergot alkaloid elymoclavine (1 mg/kg, i. p.) tends to have a positive effect on learning and memory. The muscarine cholinergic receptor blocker scopolamine (2 mg/kg, i. p.) substantially deteriorates short-term memory in passive avoidance training. Piracetam in a dose of 600 mg/kg does not change the negative effect of scopolamine on the memory, while centrophenoxine (100 mg/kg) and elymoclavine (1 mg/kg) eliminate it in view of the fact that the impairment of the short-term memory in the reported experiments was induced by the cholinolytic agent scopolamine, it should be assumed that the observed effects of piracetam, centrophenoxine and elymoclavine are due to definite interactions between the cerebral cholinergic and monoaminergic mechanisms.

Topics: Animals; Avoidance Learning; Ergolines; Glycolates; Male; Meclofenoxate; Memory, Short-Term; Piracetam; Pyrrolidinones; Rats; Rats, Inbred Strains; Scopolamine

The GABAergic influence on the effects of the ergot alkaloid elymoclavine on exploratory behaviour of rats in open field was studied using pharmacological tools. GABA was found to reduce the elymoclavine-stimulated exploratory behaviour. The GABAergic antagonists picrotoxin and bicuculline reversed the influence of GABA on the elymoclavine effects. The indirect GABA mimetics nipecotic acid, aminooxyacetic acid, di-n-propylacetate and semicarbazide also reduced the behavioural effects of elymoclavine. The data suggest some role of different functional states of the GABA system and of the close relationship between GABA and DA in the brain in the effects of elymoclavine on exploratory behaviour.

Topics: Aminooxyacetic Acid; Animals; Bicuculline; Ergolines; Exploratory Behavior; gamma-Aminobutyric Acid; Male; Nipecotic Acids; Picrotoxin; Rats; Rats, Inbred Strains; Receptors, GABA-A; Semicarbazides

Pharmacological investigations of the ergot alkaloid of the group of clavines, elymoclavine, isolated from Claviceps sp. cp. II showed the following results: The LD50 for mice for 24 h was 350 (228-535) mg/kg and for rats 145 (81-258) mg/kg. Elymoclavine induced a dose-dependent stereotypy (doses of 2 to 10 mg/kg) in rats and mice which was antagonized by haloperidol and pimozide. It prevented the development of haloperidol catalepsy in rats and produced rotations contralateral to the striatal lesions with 6-OHDA which were antagonized by pimozide and partly by cyproheptadine. Elymoclavine, like bromocriptine, decreased the plasma level of prolactin. Furthermore, elymoclavine increased the exploratory activity of rats in open field; this effect was antagonized by haloperidol and was essentially influenced by many substances acting on different transmitter systems (NA, DA, GABA). Elymoclavine inhibited the picrotoxin and electroshock convulsive seizures but potentiated the pentylenetetrazol ones in mice as these effects were differently influenced by pimozide, haloperidol, 5-HTP, atropine and phentolamine. 100 and 250 micrograms/kg of elymoclavine produced a considerable and persisting decrease of the blood pressure in anaesthetized cats. At 1 X 10(-6) M, without producing any per se effect, elyoclavine decreased the contractile effects of acetylcholine, nicotine, BaCl2 and PGE1 as well as the field electrical stimulation-induced contractions in an isolated segment from guinea-pig ileum. The observed effects of elymoclavine are mainly due to its dopaminergic agonist action. It seems, however, that influences on other transmitter receptors also underlie the mechanism of action of this ergot alkaloid.

Topics: Animals; Behavior, Animal; Blood Pressure; Catalepsy; Corpus Striatum; Ergolines; Exploratory Behavior; Guinea Pigs; Humans; Hydroxydopamines; Ileum; Lethal Dose 50; Mice; Muscle Contraction; Muscle, Smooth; Oxidopamine; Prolactin; Rats; Seizures; Stereotyped Behavior

The pharmacological investigation of the ergot alkaloid of the group of clavines elymoclavine isolated from Claviceps sp. cp. II showed the following: LD50 for mice for 24 hours was 350 (228 divided by 535) mg/kg and for rats--145 (81 divided by 258) mg/kg; elymoclavine induced a dose-dependent stereotypy in rats and mice which was antagonized by haloperidol and pimozide; it prevented the development of haloperidol catalepsy in rats and produced rotations contralateral to the striatal lesions with 6-OHDA which were antagonized by pimozide and partly by cyproheptadine. Elymoclavine increased the exploratory activity of rats in open field as this effect was antagonized by haloperidol and was essentially influenced by many substances acting on different transmitter systems (NA, DA, GABA). Elymoclavine inhibited the picroroxin and electroshock convulsive seizures but potentiated the pentylenetetrazol ones in mice as these effects were differently influenced by pimozide, haloperidol, 5-HT, atropine and phentolamine. The observed effects of elymoclavine are mainly due to its DAergic agonistic action. It seems, however, that influences on other transmitter receptors also underlie the mechanism of action of this ergot alkaloid.

Topics: Animals; Bromocriptine; Catalepsy; Central Nervous System; Corpus Striatum; Ergolines; Exploratory Behavior; Haloperidol; Humans; Hydroxydopamines; Mice; Oxidopamine; Pimozide; Rats; Receptors, Dopamine; Seizures; Stereotyped Behavior; Sympathetic Nervous System

L-Tryptophan did not exert any influence on peptide alkaloid formation in an ergotamine and in an ergosine-accumulating C. purpurea strain. A different picture was observed in a series of related C. purpurea strains. Tryptophan showed a slight stimulatory effect on the ergotoxine producer Pepty 695/S. A blocked mutant of it, designated as Pepty 695/ch which was able to accumulate secoclavines gave similar results. In a high-yielding elymoclavine strain Pepty 695/e, the progeny of the former one, tryptophan up to a concentration of 25 mM stimulated remarkably clavine biosynthesis. Furthermore, tryptophan could overcome the block of synthesis by inorganic phosphate. Increased specific activities of chanoclavine cyclase but not DMAT synthetase were observed in cultures of strain Pepty 695/e supplemented with tryptophan. 5-Methyltryptophan and bioisosteres of tryptophan were ineffective in alkaloid stimulation. These results are compared with those obtained with the grass ergot strain SD 58 and discussed with the relation to other induction phenomena.

Topics: Chemical Phenomena; Chemistry; Claviceps; Culture Media; Ergolines; Ergot Alkaloids; Mutation; Phosphates; Stereoisomerism; Tryptophan

The effect of elymoclavine on rat open field behaviour was examined. The participation of catecholaminergic mechanisms in the elymoclavine effect on rat open field behaviour was also studied using pharmacological tools for analyzing these mechanisms. Elymoclavine in increasing doses stimulated ambulation and rearing but inhibited defecation. Haloperidol blocked the responses to all doses of elymoclavine. The effect of elymoclavine on ambulation was potentiated by propranolol, desipramine and pargyline. Its effect on rearing was reduced by desipramine, pargyline, alpha-methyl-p-tyrosine (alpha-MpT), L-DOPA, DDC (diethyldithiocarbamate) and 6-OHDA (applied intracerebroventricularly). The effect of elymoclavine on defecation was reduced only by DDC. It is concluded that elymoclavine influences both locomotor components of exploratory behaviour in the open field (ambulation and rearing) and defecation. These behavioural effects of elymoclavine are realized through the participation of central catecholaminergic mechanisms (dopaminergic and to a less extent noradrenergic).

Topics: Animals; Behavior, Animal; Catecholamines; Defecation; Drug Interactions; Ergolines; Exploratory Behavior; Male; Motor Activity; Rats; Rats, Inbred Strains; Synaptic Transmission

The mechanism by which estrogens inhibit castration atrophy has been investigated morphologically and biochemically utilizing ventral prostate from Copenhagen rats. The suppression of weight loss and gross edematous appearance of the prostate associated with the in vivo effect of 17 beta-estradiol (E2) could not be accounted for by DNA and protein synthesis. Increase in the fluid content in the tissues was confirmed by demonstration of significant increase in the ratio of wet/dry tissue weights. Light microscopy demonstrated that the main effects were on the stroma, characterized by large interglandular areas almost totally devoid of collagen resulting in an edematous appearance. Electron microscope studies showed an abundance of fluid localized adjacent to the capillary endothelium and some red blood cells indicating disturbances in capillary permeability. The combination of a prolactin secretion inhibiting agent with E2 alone, indicating an involvement of prolactin the estrogen effect. Differences in blood prolactin concentration between the strains of rats may influence the sensitivity of the prostate to estrogens.

Topics: Animals; Castration; DNA; Ergolines; Estradiol; Estrogens; Male; Organ Culture Techniques; Organ Size; Prolactin; Prostate; Proteins; Rats; Rats, Inbred Strains

A new synthesis is described for 10alpha-methoxy-delta8,9-lysergaldehyde involving the oxidation of elymoclavine with manganese dioxide in methanol. Lysergol and agroclavine provide no reaction under the same conditions.

Topics: Ergolines; Lysergic Acid; Manganese; Methods; Oxidation-Reduction

Topics: Abnormalities, Drug-Induced; Animals; Ergolines; Ergot Alkaloids; Female; Fetal Death; Fetal Resorption; Hot Temperature; Lumbar Vertebrae; Male; Mice; Pregnancy; Pregnancy Complications; Ribs; Stress, Physiological; Teratogens; Thoracic Vertebrae

Topics: Cardiovascular Agents; Central Nervous System Stimulants; Ergolines; Ergot Alkaloids; Oxytocics; Reserpine