Cholangiocarcinoma (CC) remains one of the most challenging biliary tract malignancies, with limited therapeutic options and poor survival rates. We report the discovery and mechanistic investigation of usnic acid (UA) enantiomers as novel anti-CC agents. We identified the lichen Cladonia foliacea as a potential source of anticancer agents and developed a sustainable protocol to isolate (S)-UA as the most abundant metabolite. Our comprehensive comparative study of both enantiomers revealed time-dependent enantio-preference in their anti-cancer activity. While (S)-UA demonstrated twice the potency at 24 h, (R)-UA exhibited nearly ten-fold greater activity at 48 h and 72 h, particularly at lower concentrations (2.9 and 29 mM). Overall, both enantiomers inhibited EGI-1 cell proliferation in the micromolar range in a dose- and time-dependent manner. In silico studies and kinase profiling identified MNK2 as a primary target, with subsequent validation confirming direct binding and inhibition. Mechanistic studies demonstrated that UA enantiomers modulate the MAPK pathway, leading to decreased phosphorylation of eIF4E and suppression of cancer-promoting proteins. The successful translation of activity from 2D cell cultures to patient-derived 3D organoid models further validates their therapeutic potential. Our findings establish usnic acid as a promising natural product scaffold for CC treatment and provide detailed insights into its mechanism of action through MNK2 targeting and MAPK pathway modulation, with important considerations for enantiomer-specific temporal efficacy.

Premise: Leaf extracts are valuable sources of bioactive compounds. However, co‐extracted chlorophylls interfere with analyses, including spectroscopic and biochemical assays. Existing methods for chlorophyll removal often have limitations, including the use of hazardous solvents, low specificity, or high costs.Methods and Results: A solid‐phase extraction protocol for chlorophyll removal from leaf extracts of Corylus avellana was developed using commercially available cartridges. The method requires standard equipment, can be completed within 10 minutes, and is scalable from analytical to preparative quantities. We validated this protocol across 20 taxa, demonstrating the removal of 85% of chlorophylls, successful scale up of quantities, cartridge reusability, and low solvent consumption.Conclusions: Key innovations of the protocol include simplified elution steps and the possibility of multiple reuse cycles. The simplicity, sustainability, and scalability of this new protocol make it particularly valuable for high‐throughput applications and process development.

Apricot kernels (Prunus armeniaca L.) represent a valuable by-product of stone fruit cultivation, offering diverse applications in food, cosmetic, and pharmaceutical industries. While apricot kernel oil is recognized for its rich composition of unsaturated fatty acids, phenolics, and tocopherols, its therapeutic potential, particularly in cancer prevention, remains unexplored. This study investigated a purified fraction (FOPF) obtained from Farclo variety kernel oil, cultivated in the Emilia-Romagna region of Italy and selected for its naturally low amygdalin content. In vitro studies demonstrated FOPF’s significant antiproliferative effects against colorectal cancer (LoVo, HT29) and hepatocarcinoma (Hep3B) cell lines, with GI₅₀ values ranging from 0.06 to 0.09 mg/mL. The fraction induced cell cycle arrest and significantly inhibited cancer cell migration, effects mediated through PPAR-γ expression modulation. These findings establish FOPF’s potential as a natural chemopreventive agent and provide a foundation for its development as a nutraceutical ingredient targeting colorectal and hepatic cancers.

Neurodegenerative diseases represent a growing global health burden. With current treatments limited to symptom management, natural biodiversity offers a promising source of novel, multi-target compounds. Bioprospecting thus emerges as a sustainable strategy for discovering innovative neuroprotective therapies, underscoring the importance of preventing biodiversity for future drug discovery. An initial metabolomic screening of Italian plants was performed to support a bioprospective approach, in which 27 plant extracts were evaluated for their ability to inhibit acetylcholinesterase and tyrosinase, targeting mechanisms involved in neurodegenerative diseases. Adenophora liliifolia and Carissa macrocarpa emerged as the most promising species through multi-parameter screening. Metabolomic analysis showed that most identified metabolites exhibit activity against acetylcholinesterase and/or tyrosinase, supporting observed bioactivities. These extracts represent potential candidates for developing therapies against neurodegenerative diseases. These findings highlight systematic bioprospecting as a powerful strategy for identifying novel candidates with neuroprotective activity. By integrating traditional knowledge with modern metabolomic and biochemical approaches, this study demonstrates how underexplored plant biodiversity, such as Adenophora liliifolia and Carissa macrocarpa, can yield promising therapeutic candidates for targeting the complex multi-target mechanisms underlying neurodegenerative disorders.

Life expectancy in high-income countries is increasing, leading to a higher incidence of age-related neurodegenerative diseases. To address this urgent medical need, several molecular targets have been identified, including advanced glycation end products (AGEs) and tyrosinase. Given the well-established role of diet in counteracting degenerative processes, this study aimed to identify a potential food ingredient with combined anti-tyrosinase and anti-glycative properties. Acanthus mollis L. was selected based on its inclusion in the BelFrIt list and its known content of tyrosinase inhibitors, such as benzoxazinones and verbascoside. Extraction of A. mollis leaves was optimized using a design of experiments approach, comparing microwave- and ultrasound-assisted techniques. Optimal conditions were achieved using microwave-assisted extraction with ethanol 80%, 80 °C, one cycle, drug-to-solvent ratio of 10 mL/g. The optimized extract (at 5 mg/mL) inhibited tyrosinase activity by approximately 47%, increasing to 58% after chlorophyll removal. Moreover, the extract reduces AGEs formation in presence of methylglyoxal, with an activity at 1 mg/mL comparable with that of a well-known anti-glycative agent. A similar trend was observed in the reduction in methylglyoxal and glyoxal levels. Overall, these results support the potential of the optimized A. mollis extract as a functional food ingredient to counteract aging-related neurodegeneration.