Laurus nobilis L. is commonly used in folk medicine in the form of infusion or decoction to treat gastrointestinal diseases and flatulence as a carminative, antiseptic, and anti-inflammatory agent. In this study, the essential oil (EO) composition of wild-grown L. nobilis L. leaves collected from seven different altitudinal locations in the Molise region and adjacent regions (Abruzzo and Campania) was investigated. EOs from the leaves were obtained by hydrodistillation and analyzed by GC-FID and GC/MS, and 78 compounds were identified. The major oil components were 1,8-cineol (43.52–31.31%), methyl-eugenol (14.96–4.07%), α-terpinyl acetate (13.00–8.51%), linalool (11.72–1.08%), sabinene (10.57–4.85%), α-pinene (7.41–3.61%), eugenol (4.12–1.97%), and terpinen-4-ol (2.33–1.25%). Chemometric techniques have been applied to compare the chemical composition. To shed light on the nutraceutical properties of the main hydrophobic secondary metabolites (≥1.0%) of laurel EOs, we assessed the in vitro antioxidant activities based on 2,2-diphenyl-1-picrylhydrazyl (DPPH•) radical scavenging activity and the reducing antioxidant power by using a ferric reducing power (FRAP) assay. Furthermore, we highlighted the anti-inflammatory effects of seven EOs able to interfere with the enzyme soluble epoxide hydrolase (sEH), a key enzyme in the arachidonic acid cascade, in concentrations ranging from 16.5 ± 4.3 to 8062.3 ± 580.9 mg/mL. Thanks to in silico studies, we investigated and rationalized the observed anti-inflammatory properties, ascribing the inhibitory activity toward the disclosed target to the most abundant volatile phytochemicals (≥1.0%) of seven EOs.

Mass spectrometry-based chemical proteomic approaches using limited proteolysis have become a powerful tool for the identification and analysis of the interactions between a small molecule (SM) and its protein target(s). Gracilioether A (GeA) is a polyketide isolated from a marine sponge, for which we aimed to trace the interactome using this strategy. DARTS (Drug Affinity Responsive Target Stability) and t-LiP-MS (targeted-Limited Proteolysis-Mass Spectrometry) represented the main techniques used in this study. DARTS was applied on HeLa cell lysate for the identification of the GeA target proteins, and t-LiP-MS was employed to investigate the protein’s regions involved in the binding with GeA. The results were complemented through the use of binding studies using Surface Plasmon Resonance (SPR) and in silico molecular docking experiments. Ubiquitin carboxyl-terminal hydrolase 5 (USP5) was identified as a promising target of GeA, and the interaction profile of the USP5-GeA complex was explained. USP5 is an enzyme involved in the pathway of protein metabolism through the disassembly of the polyubiquitin chains on degraded proteins into ubiquitin monomers. This activity is connected to different cellular functions concerning the maintenance of chromatin structure and receptors and the degradation of abnormal proteins and cancerogenic progression. On this basis, this structural information opens the way to following studies focused on the definition of the biological potential of Gracilioether A and the rational development of novel USP5 inhibitors based on a new structural skeleton.

Vicia faba L. is a widely cultivated legume known to contain numerous specialised metabolites. In this study, the seed coats and cotyledons of two ancient V. faba L. varieties, historically consumed in southern Italy and distinguished by black and purple seed coats, were extracted using 80% methanol and 80% ethanol. Extracts were analysed for total polyphenol, flavonoid and proanthocyanidin contents, and antioxidant activity using DPPH, ABTS, and FRAP assays. The purple seed coats exhibited the highest levels of phenolics and antioxidant capacity, exceeding those of black seed coats. Next, liquid chromatography coupled with high-resolution mass spectrometry (LC-ESI-HR-MS) was used to characterise the bioactive metabolites in both seed coats and cotyledons. The purple variety showed a higher phytochemical content, with a greater level of flavonoids and proanthocyanidins in methanolic extract. Furthermore, the purple seed coat exhibited in vitro anti-inflammatory activity by inhibiting soluble epoxide hydrolase (sEH), a key enzyme in the arachidonic acid cascade, with an IC₅₀ of 31.51 ± 1.16 µg/µL. Elemental analysis was performed for both varieties to assess their nutritional value. Specifically, the purple seed coats were found to represent a valuable source of bioactive compounds and micronutrients, highlighting their potential applications in nutraceutical, cosmetic, and food supplement sectors.

The intrinsically disordered protein (IDP) c-Myc plays a crucial role in regulating cell growth, proliferation, and apoptosis, making it a significant target for cancer therapy. Herein, we report an integrated approach combining computational and biological methods to uncover key c-Myc structural aspects aimed at identifying modulators of this protein. A virtual screening campaign starting from commercially available libraries was performed, leading to the selection of 11 promising c-Myc binders. Preliminary biophysical and biological analysis, including Surface Plasmon Resonance (SPR) and MTT [(3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide)] assays, highlighted three new potential c-Myc/Max heterodimerization inhibitors. Among them, 11 emerged as the most active one, displaying promising dissociation costant (K_D) values on c-Myc (~ 60 μM), only slightly higher than the known binder 10058-F4 (with a K_D ~ 40 μM) and on Max (with a K_D ~ 80 μM). Notably, 11 showed significant antiproliferative effects on three different malignant cell lines (i.e., U937, HCT-116, and HT-29, as models of acute myeloid leukemia and colorectal cancer cell lines, respectively) in a dose- and cell-dependent manner. Importantly, a significant decrease in c-Myc levels was observed, especially in U937 leukemia cells with an 85 % reduction. Finally, molecular dynamics simulations and proteomic studies were performed to identify its binding site and key interacting residues. In summary, this work presents a robust research strategy and offers valuable insights for identifying new c-Myc/Max disruptors.

Wild plant biodiversity represents a largely untapped source of chemically diverse metabolites. However, many wild edible species remain poorly characterized despite their potential as sources of antioxidant and anti-inflammatory phytochemicals. Daucus carota L. subsp. carota, commonly known as wild carrot, has attracted interest due to its long history of medicinal use. In this study, seed extracts collected from populations growing in two Italian regions (Lazio and Molise) were investigated to evaluate their phytochemical composition, mineral content, antioxidant activity, and anti-inflammatory potential. The metabolite profiles were characterized by qualitative Liquid Chromatography-Mass Spectrometry analysis, allowing the identification of several phenolic compounds and other secondary metabolites. Antioxidant activity was assessed using in vitro assays, while anti-inflammatory activity was evaluated through the inhibition of cyclooxygenase and soluble epoxide hydrolase, two key enzymes involved in the arachidonic acid cascade. To gain further insight into the possible mechanisms underlying these activities, molecular docking analyses were performed on selected metabolites identified in the extracts, exploring their interactions with the target enzymes. The extracts displayed promising antioxidant and anti-inflammatory properties, while docking results supported the potential role of specific metabolites in enzyme modulation. These findings highlight the value of underexplored wild D. carota seeds as a source of natural compounds with potential applications in the development of nutraceutical and cosmeceutical products.