Ethyl acetate extract from Dinizia excelsa Ducke wood: A potential phytotherapeutic agent with antioxidant, cytotoxic and hemolytic (in normal cells), immunomodulatory, antimicrobial, antiparasitic and antitumor activities

Amazonian biodiversity is an underexploited source of bioactive natural products with potential relevance to drug discovery for neglected diseases. Here, we characterized the chemistry and multi-target in vitro activity of an ethyl acetate extract from Dinizia excelsa wood. Extraction yielded 4.84 ± 0.10% and produced a phenolic-rich fraction with high total phenolics (501.8 ± 1.0 mg GAE/g extract), flavonoids (172.4 ± 0.8 mg QE/g), flavonols (49.7 ± 0.3 mg QE/g), and hydrolyzable tannins (229.4 ± 1.2 mg TAE/g). LC-HRMS dereplication annotated 19 metabolites, dominated by glycosylated flavonoids (rutin, isoquercetin), phenolic acids (ferulic and ellagic acids), ellagic derivatives (urolithins), and galloylated hydrolyzable tannins. The extract showed moderate-to-low antioxidant capacity across complementary assays (IC₅₀ 78.38-161.99 µg/mL) and a favorable safety profile, with moderate/low cytotoxicity in mammalian cell lines (IC₅₀ 364-700 µg/mL) and minimal hemolysis (<6% up to 1000 µg/mL). In murine splenocytes, the extract preserved membrane integrity, induced dose-dependent proliferation, and promoted a broad but controlled cytokine response (↑IL-2, IL-4, IL-10, IL-12, IFN-γ, IL-17, TNF-α; regulated TGF-β), alongside reduced NO, mild cytosolic ROS modulation, and stable mitochondrial membrane potential. Notably, it displayed strong antiparasitic activity against Trypanosoma cruzi (IC₅₀ 2.37-3.94 µg/mL) and Leishmania spp. (IC₅₀ 1.83-6.43 µg/mL), with additional activity against Schistosoma mansoni (IC₅₀ 114.83-247.0 µg/mL) and Plasmodium falciparum 3D7 (IC₅₀ 5438.27 ng/mL). Antibacterial MICs ranged 16-512 µg/mL and antifungal MICs 16-256 µg/mL, consistent with bacteriostatic/fungistatic effects.