Identification and Characterization of Novel Components in Psilocybin-Containing Mushroom Extracts and Their Effects on Inflammation-Related Disorders

Tryptamine producing fungi, and especially their major metabolite psilocybin, have been revisited in
recent years as potential pharmaceutical agents for the alleviation of various disorders. Psilocybin’s
effects are mediated mainly via the 5-HT (serotonin) receptor family, which is abundant in all major
human organ systems. Specifically, the enteric nervous system is a producer and consumer of 95% of
5-HT. In addition to psilocybin, psychedelic mushrooms produce many other tryptamine class
compounds that have yet to be characterized but believed to possess high therapeutic potential. To
date, there are over 140 known psilocybin-Containing Mushroom species worldwide, all of which
exhibit different secondary metabolite profiles. Previous studies implemented various methods of
extraction and quantification to assess these compounds but culminated in high variance results.
Here, we have developed an optimal extraction method for the tryptamine fraction from fruiting
bodies, successfully quantified these metabolites via LCMS/MS and managed to dissolve the fungal
matrices in a non-toxic hydrophilic solvent to create extracts suitable for in-vitro experiments.
Furthermore, to test the active metabolites from fungi, we standardized the in-vitro
dephosphorylation of psilocybin to its active metabolite psilocin using alkaline phosphatase. We
chose THP1 monocyte human cell line as the model to evaluate the immunological effects of
different fungi extracts. We succeeded to cultivate, differentiate and polarize THP1 cells to pro-
inflammatory macrophages. In addition, quantitative RT-PCR scanning of THP1 showed high
expression of 5-HT7 and 5-HT2b receptors, to which psilocin is highly affine. This study will reveal
and characterize novel secondary metabolites that naturally occur in fungi and help assert their
effects and therapeutic potential.