Free radicals play a critical role in the chemical processes that occur in all cells. Pharmaceutical companies manufacture a variety of synthetically prepared antioxidants, but it is known that many of these can be carcinogenic. As a result, efforts are being made to find natural antioxidants that do not have these side effects. Lichens may be suitable candidates because they contain secondary metabolites with proven antioxidant properties. This could be explained by the presence of compounds with phenolic groups in lichens. The radical scavenging reaction is a chemical reaction governed by stoichiometry, and our aim is to determine the efficacy of these reactions. The aim of this study is to compare metabolite activity based on the same amount of substance involved in radical scavenging, calculated in micromoles rather than weight concentration. This provides an accurate way of comparing radical scavenging activity. We tested superoxide anion scavenging activity and free radical scavenging activity of isolated lichen secondary metabolites and their mixtures in different ratios. The following compounds were isolated and tested for antioxidant activity: gyrophoric acid (Umbilicaria hirsuta), evernic acid (Evernia prunastri), physodic acid, 3-hydroxyphysodic acid, physodalic acid and atranorin (Hypogymnia physodes), and usnic acid (as a synthetic compound). Of all the tested compounds, 3-hydroxyphysodic acid, as well as mixtures containing this metabolite, showed the strongest scavenging activity. The results also demonstrated that calculation by amount of substance leads to a new consideration of antioxidant activity.

Lichen metabolites represent a wide range of substances with a variety of biological effects. The present study was designed to analyze the potential antiproliferative, antimicrobial and antioxidative effects of several extracts from lichens (Pseudevernia furfuracea, Lobaria pulmonaria, Cetraria islandica, Evernia prunastri, Stereocaulon tomentosum, Xanthoria elegans and Umbilicaria hirsuta) and their secondary metabolites (atranorin, physodic acid, evernic acid and gyrophoric acid). The crude extract, as well as the isolated metabolites, showed potent antiproliferative, cytotoxic activity on a broad range of cancer cell lines in 2D (monolayer) and 3D (spheroid) models. Furthermore, antioxidant (2,2-diphenyl-1-picryl-hydrazylhydrate (DPPH) and in vitro antimicrobial activities were assessed. Data showed that the lichen extracts, as well as the compounds present, possessed biological potential in the studied assays. It was also observed that the extracts were more efficient and their major compounds showed strong effects as antiproliferative, antioxidant and antibacterial agents. Moreover, we demonstrated the 2D and 3D models’ importance to drug discovery for further in vivo studies. Despite the fact that lichen compounds have been neglected by the scientific community for long periods, nowadays they are objects of investigation based on their promising effects.