Ergot alkaloids: synthetic approaches to lysergic acid and clavine alkaloids

Ergot alkaloids are among the most important pharmaceuticals and natural toxins.

Total synthesis, biosynthesis and biological profiles of clavine alkaloids

This review highlights noteworthy synthetic and biological aspects of the clavine subfamily of ergot alkaloids.

Alkaloid Cluster Gene ccsA of the Ergot Fungus Claviceps purpurea Encodes Chanoclavine I Synthase, a Flavin Adenine Dinucleotide-Containing Oxidoreductase Mediating the Transformation of N-Methyl-Dimethylallyltryptophan to Chanoclavine I

ABSTRACT Ergot alkaloids are indole-derived secondary metabolites synthesized by the phytopathogenic ascomycete Claviceps purpurea. In wild-type strains, they are exclusively produced in the sclerotium, a hibernation structure; for biotechnological applications, submerse production strains have been generated by mutagenesis. It was shown previously that the enzymes specific for alkaloid biosynthesis are encoded by a gene cluster of 68.5 kb. This ergot alkaloid cluster consists of 14 genes coregulated and expressed under alkaloid-producing conditions. Although the role of some of the cluster genes in alkaloid biosynthesis could be confirmed by a targeted knockout approach, further functional analyses are needed, especially concerning the early pathway-specific steps up to the production of clavine alkaloids. Therefore, the gene ccsA, originally named easE and preliminarily annotated as coding for a flavin adenine dinucleotide-containing oxidoreductase, was deleted in the C. purpurea strain P1, which is able to synthesize ergot alkaloids in axenic culture. Five independent knockout mutants were analyzed with regard to alkaloid-producing capability. Thin-layer chromatography (TLC), ultrapressure liquid chromatography (UPLC), and mass spectrometry (MS) analyses revealed accumulation of N-methyl-dimethylallyltryptophan (Me-DMAT) and traces of dimethylallyltryptophan (DMAT), the first pathway-specific intermediate. Since other alkaloid intermediates could not be detected, we conclude that deletion of ccsA led to a block in alkaloid biosynthesis beyond Me-DMAT formation. Complementation with a ccsA/gfp fusion construct restored alkaloid biosynthesis. These data indicate that ccsA encodes the chanoclavine I synthase or a component thereof catalyzing the conversion of N-methyl-dimethylallyltryptophan to chanoclavine I.

Ergot alkaloids: extent of human and animal exposure

Ergot alkaloids are formed by Claviceps spp. on grains and grasses and by fungal endophytes such as Neotyphodium spp. in grasses, notably tall fescue and perennial ryegrass. Ergots from grains and grasses show a wide variation in alkaloid composition. The main ergot alkaloids are pharmacologically active lysergic acid derivatives – e.g. ergometrine (ergonovine), ergotamine, ergosine, ergocornine, α-ergocryptine, ergocristine, and ergovaline; derivatives of isolysergic acid, e.g. ergotaminine; and clavine alkaloids, e.g. agroclavine. Other structurally unrelated toxic alkaloids such as lolitrems are formed by fungal endophytes in grasses. The present review focuses more on how man and animals are exposed to ergot alkaloids than on toxicology and methods of analysis. Ergot poisoning in humans, well known in the Middle Ages, can be of two types: convulsive ergotism and gangrenous ergotism. Since the beginning of the last century there have been outbreaks in Russia, England, India, France and Ethiopia. The principal route of human exposure to ergot alkaloids is by consumption of contaminated food; another route is inhalation of grain dust. Toxicoses in animals due to ergot alkaloids are more common, particularly poisoning of livestock grazing on endophyte infected grasses. Analyses in Canada, Germany, Switzerland, Sweden and Denmark found ergot alkaloids in human foods such as wheat and rye flours, bread, and other grain foods, often at levels greater than 1000 µg/kg. Processing studies have confirmed that the alkaloids survive baking; they also remain to some extent after brewing of beer. There is little evidence for carryover of ergot alkaloids into animal tissue and milk. As an indication of the importance of controlling ergot for the health of animals and people, Canada, the European Union, Switzerland, USA, Japan, Australia and New Zealand have regulations for ergot in grains but only Uruguay and Canada have regulations for the actual ergot alkaloids in feed.

N-1-Trimethylsilyl Derivatives of Ergot Alkaloids

N-1-Trimethylsilyl derivatives of five different suitably protected parent ergot (clavine) alkaloids (agroclavine 1a, elymoclavine 2a, lysergol 3a, lysergene 4a, and 9,10-dihydrolysergol 5a) were prepared in 47-94% yields by refluxing the (protected) parent compounds with N-methyl-N-(trimethylsilyl)trifluoroacetamide in acetonitrile under nitrogen atmosphere.