Production of Cholinesterase-Inhibiting Compounds in In Vitro Cultures of Club Mosses

  • Wojciech J. SzypułaEmail author
  • Agnieszka Pietrosiuk
Living reference work entry
Part of the Reference Series in Phytochemistry book series (RSP)


This review provides a comprehensive overview of the cholinesterase-inhibiting compound production in in vitro culture of club mosses (Lycopodiaceae sensu lato). Some hallmarks in a rich history of studies on club mosses, their complex systematics and phylogeny, and secondary metabolites with potential medicinal uses are presented. The review summarizes the published literature, from historical times to the present, including reports on the latest developments in the biosynthesis of alkaloids using different methods. Currently, over 30 compounds which are AChE inhibitors are being assessed in various phases of preclinical and clinical trials, and a few have been approved for use. Huperzine A (HupA, selagine), an alkaloid isolated from some club mosses, has been chosen as a promising drug candidate for Alzheimer’s disease. Although the procedure for the total HupA synthesis has been developed, the pharmaceutical industry uses mainly Huperzia serrata sporophytes collected in its natural habitat. Currently, a total of approximately 350 different Lycopodium alkaloids are known, and some of them demonstrate very strong acetylcholinesterase inhibitory or butyrylcholinesterase-inhibiting activity. Since the 1950s, a number of approaches to Lycopod regeneration have been described, and some appeared to be potentially useful for large-scale propagation of the plant material, including the production of secondary metabolites. Some other established cultures which partially succeeded used Lycopod gametophytes or sporophytes. Some of those studies dealt specifically with the biosynthesis of alkaloids and obtaining the biomass for their isolation. These values clearly demonstrate that the highest HupA content in the plant material from in vitro tissue cultures exceeds by approximately 26-fold or by 13-fold the mean HupA level in the whole plant of H. serrata or by 10-fold the highest HupA content in the H. serrata sporophytes with the most efficient biosynthesis of the alkaloid. Importantly, the in vitro methods successfully shorten the life cycle of the plants which under natural conditions can take many years to develop.


Club mosses Huperzine A Lycopodiaceae Huperziaceae Alkaloids Acetylcholinesterase inhibitory In vitro culture 



Murashige and Skoog medium with half strength mineral salt content and full strength of organic component






Alzheimer’s disease




Central nervous system


Huperzine A (selagine)


Huperzine B


Sensu lato


Sensu stricto


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Authors and Affiliations

  1. 1.Department of Pharmaceutical Biology and Medicinal Plants Biotechnology, Faculty of PharmacyThe Medical University of WarsawWarsawPoland

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