The effects of ergot and non-ergot-derived dopamine agonists in an experimental mouse model of endometriosis.

Implantation of a retrogradely shed endometrium during menstruation requires an adequate blood supply, which allows the growth of endometriotic lesions. This suggests that the development of endometriosis can be impaired by inhibiting angiogenesis. The growth of endometriotic foci is impaired by commercial oncological antiangiogenic drugs used to block vascular endothelial growth factor (VEGF) signaling. The dopamine agonist cabergoline (Cb2) inhibits the growth of established endometriosis lesions by exerting antiangiogenic effects through VEGFR2 inactivation. However, the use of ergot-derived Cb2 is associated with an increased incidence of cardiac valve regurgitation. To evaluate the potential usage of non-ergot-derived dopamine agonists for the treatment of human endometriosis, we compared the efficacy of quinagolide with that of Cb2 in preventing angiogenesis and vascularization in a heterologous mouse model of endometriosis. Nude mice whose peritoneum had been implanted with eutopic human endometrial fragments were treated with vehicle, 50  µg/kg per day oral Cb2, or 50 or 200  µg/kg per day quinagolide during a 14-day period. At the end of the treatment period, the implants were excised in order to assess lesion size, cell proliferation, degree of vascularization, and angiogenic gene expression. Neoangiogenesis was inhibited and the size of active endometriotic lesions, cellular proliferation index, and angiogenic gene expression were significantly reduced by both dopamine agonists when compared with the placebo. Given that Cb2 and quinagolide were equally effective in inhibiting angiogenesis and reducing lesion size, these experiments provide the rationale for pilot studies to explore the use of non-ergot-derived dopamine agonists for the treatment of endometriosis in humans.

Liquid chromatography-tandem mass spectrometry characterization of ergocristam degradation products.

The UPLC method with diode array UV detection was developed for qualitative determination of ergocristine and ergocristam including degradation products. The mechanism of the ergocristam disruptive reaction was described based on MS/MS characterization of ammonolytic product, N-(d-lysergyl)-l-valinamide (A1) and two methanolytic products, methyl ester of N-(d-lysergyl)-l-valine (M2), and N-[N-(d-lysergyl)-l-valyl]-l-phenylalanyl-d-prolyl methyl ester (M1). The influence of extraction conditions on epimerization and degradation of ergocristine and ergocristam was tested and conditions for reproducible decomposition of ergocristam were found. The presented method could potentially be applied for ergot alkaloids determination in sclerotia, fermentation broth, mycelium, and possibly contaminated food products, i.e. corn, flour, bread, etc., and feeding stuffs containing ungrounded cereals.

A novel ergot alkaloid as a 5-HT(1A) inhibitor produced by Dicyma sp.

In the course of a search for small-molecule inhibitors of 5-hydroxytryptamine receptors we have identified a novel ergoline derivative (1) from the fungal culture of Dicyma sp. This compound has a pK(i) of 10.2 versus the 5-hydroxytryptamine(1A) receptor subtype. The structure was elucidated by extensive NMR spectroscopy and mass spectrometry.

Better biosynthesis of ergot alkaloids.

Different culture media containing different effectors for production of ergot-alkaloids were prepared. The culture media were inoculated with equal mycelia of Claviceps purpurea and some were incubated for 22 days and some incubated for 45 days. The medium that contained a mixture of glucose, glycine and tryptophan showed highest yield of the alkaloids and reached the maximum period for production of agroclavine faster.

Ergot alkaloids produced by submerged cultures of Claviceps zizaniae.

Two ergopeptine alkaloids, alpha-ergocryptine (1) and its C(8) epimer alpha-ergocryptinine, have been isolated from the mycelium and fermentation broth of submerged cultures of Claviceps zizaniae CCM 8240. The structure of 1 was determined by HPLC/positive ion APCI MS and NMR analysis. Alkaloid concentrations of 10 microg/mL in 14-day-old fermentation broth and 1 mg/g of dry mycelium mass were found. These results are of considerable biotechnological interest since these were the only detectable alkaloids produced. Toxicity of naturally occurring sclerotia of C. zizaniae cannot be excluded.

Direct resolution of optically active isomers on chiral packings containing ergoline skeleton. 6. Enantioseparation of profens.

The enantioselective behaviour of some underivatized 2-arylpropionic acids (profens) and flobufen by HPLC using a terguride-based chiral stationary phase was tested. X-ray analysis of crystals of the chiral selector and its complexes with naproxen allowed a deeper insight into the enantiodiscriminative process. The column stability and reproducibility, and the potential of the packing for semipreparative scale separations were also determined. A method for determining flobufen enantiomers and metabolites in plasma samples is described.

1-Methoxy-agroclavine from Penicillium sp. WC75209, a novel inhibitor of the Lck tyrosine kinase.

A high-throughput screen was developed and implemented to identify inhibitors of the Lck tyrosine kinase. This report describes the identification of a specific inhibitor of this enzyme from the solid fermentation culture of the Penicillium sp., WC75209. The active compound was isolated and structurally characterized as 1-methoxy-5R, 10S-agroclavine, a new member of the ergot alkaloid family.

Genetics of ergoline alkaloid formation in Penicillium roquefortii.

Auxotrophic, spore color, and alkaloid biosynthetic mutants of Penicillium roquefortii were selected after N-methyl-N'-nitro-N-nitrosoguanidine treatment. Diploids were obtained via protoplast fusion techniques, and the segregants from a diploid were genetically analyzed. The data demonstrated the potential of parasexual recombination in this organism. Evidence was obtained which suggests that the his and sts (sensitivity to Sulfatase) genes may be linked. The genetic information obtained in this study can serve as a starting point for further mapping of genes in P. roquefortii, and indications are that this organism may serve as a promising vehicle for the genetic study of the formation of ergoline alkaloids.

[Mutagenicity testing of commercially used strains of P. camemberti and P. roqueforti]. / Mutagenitätsprüfung industriell verwendeter P. camemberti- und P. roqueforti-Stämme.

We tested the mutagenic potential of crude extracts of 18 strains of Penicillium camemberti and 6 strains of P. roqueforti, which are used commercially in the production of mould ripened cheese in Switzerland. No mutagenic activity could be detected in any of the extracts. Roquefortine, a mycotoxin of P. roqueforti, often found in Blue cheese, was negative in the Amestest. The results obtained do not lead reasons for experting undesired long term effects from the consumption of mould ripened cheese.

Production of fumigaclavine A by Aspergillus tamarii Kita.

Aspergillus tamarii Kita. isolated from seeds of Paspalum scrobiculatum L. is found to produce ergot alkaloids in submerged culture. The culture filtrate and mycelium are observed to contain 0.125 mg/mL and 1.2 mg/g (dry weight) total alkaloids consisting of 86.5 and 91.3% fumigaclavine A, respectively. The identification of the compound was confirmed by high-performance liquid chromatography, ultraviolet, infrared, and mass spectrophotometry analyses. This is the first report of the production of ergot alkaloid by this fungus. The possible role of fumigaclavine A as a mycotoxin is discussed.

Novel metabolites from Penicillium crustosum, including penitrem E, a tremorgenic mycotoxin.

Two new indolic metabolites were isolated from Penicillium crustosum and separated from other penitrem mycotoxins by high-performance liquid chromatography. Penitrem D is a deoxy-penitrem A. Penitrem E is dechloro-penitrem A and was shown to be tremorgenic in mice, although it has only one-third of the activity of penitrem A. Roquefortine was also shown, for the first time, to be an important metabolic product of P. crustosum.

[Lysergic acids. II. Isolation and separation of lysergic acids (author's transl)]. / Lysergsäure II. Isolation und Trennung der Lysergsäuren

During studies on the isolation of both lysergic acids, D-and D-iso-, from hydrolytic mixtures of ergot alkaloids, it became necessary to find a simple chromatographic system for column isolation of lysergic acids. The column with controlled pore glass is highly effective for these purposes. The separation of both isomeric lysergic acids occurs on the column with silica gel. The assay and the composition of lysergic acid are estimated by thin-layer chromatography on precoated plates.