Eco-Friendly Species <i>Evernia prunastri</i> (L.) Ach.: Phenolic Profile, Antioxidant, Anti-inflammatory, and Anticancer Properties

Growing in prominence is the utilization of natural product-based therapies, especially edibles used in traditional medicine, as more people seek natural and holistic approaches to health and well-being. This research aimed to determine the phenolic compounds, and antioxidant, anti-inflammatory, and anticancer effects of aqueous and methanolic extracts from <i>Evernia prunastri</i> (L.) Ach., a common spice in Algeria. HPLC-DAD was used to establish the phenolic profile, whereas the antioxidant activity was determined by DPPH, FRAP, phosphomolybdate, and hydrogen peroxide (H₂O₂) assays. Human red blood cell (HRBC) stabilization, albumin denaturation, and proteinase inhibition procedures were performed to investigate the anti-inflammatory activities, and an MTT assay was used to demonstrate the cytotoxic effect against three human cancer cell lines (HT-29, PC-3, A549) and human nontumor (CCD18-Co) cells. Our results showed that the major phenolics detected were mostly phenylpropanoids with domination of rosmarinic acid (79.17 mg/g), caffeic acid (46.52 mg/g), <i>trans</i>-cinnamic acid (29.23 mg/g), and chlorogenic acid (23.68 mg/g). In addition, six flavonoids were identified (1.98-11.34 mg/g), namely, luteolin, myricetin, kaempferol, rutin, apigenin, and quercetin. Other compounds were relatively present in both extracts, gallic acid and 3-hydroxybenzoic acid (phenolic acids), except pyrocatechol (benzenediol), which was slightly detected in the aqueous extract (0.91 mg/g). Moreover, both <i>E. prunastri</i> extracts showed strong antioxidant activity, with a higher antioxidant potential, as shown by the methanolic extract. Likewise, both reduced HRBC hemolysis damage and moderately suppressed protein denaturation, which reflected their anti-inflammatory potential. Interestingly, the methanolic extract significantly reduced the growth of HT-29, PC-3, and A549 cells by 67.03, 75.56, and 62.96% respectively. No cytotoxic effects were observed in the nontumor cells. The methanolic extract had the lowest IC₅₀ values of 100 ± 0.04, 146 ± 0.05, and 112 ± 0.06 μg/mL against HT-29, PC-3, and A549 cell lines, respectively. In conclusion, <i>E. prunastri</i>, especially its methanolic extract, could be considered as a promising source of antioxidant and anticancer molecules.