Search Results (481)

Mycobacterium avium subsp. paratuberculosis (MAP) causes Johne’s disease in ruminants and is difficult to control once persistent infection is established. We designed a MAP multi-epitope vaccine candidate by screening conserved proteins and assembling predicted B-cell, MHC class I-restricted cytotoxic T lymphocyte (CTL), and MHC class II-restricted helper T lymphocyte (HTL) epitopes into a recombinant construct. The construct was examined by physicochemical analysis, structural modeling, molecular docking, and immune simulation, then expressed in Escherichia coli and tested in BALB/c mice as a recombinant protein without external adjuvant or as an exploratory MYY-containing formulation, alongside PBS and inactivated-MAP controls. RAW264.7 macrophages were stimulated with the purified recombinant protein for RNA sequencing. Seven proteins were retained as antigen sources, and the final construct contained seven HTL epitopes, five B-cell epitopes, and ten CTL epitopes. Immunized mice produced antigen-specific IgG, IgG1, and IgG2a, and antigen-restimulated splenocytes secreted higher levels of IFN-γ, IL-2, TNF-α, and IL-17. Vaccine-induced antibodies recognized both the recombinant protein and components present in MAP lysate while showing limited reactivity toward the bovine tuberculosis diagnostic antigen PPD, and sequence analysis predicted no substantial similarity between the construct and the diagnostic antigens ESAT-6, CFP-10, MPB70, and MPB83, supporting potential DIVA compatibility. In macrophages, recombinant-protein stimulation changed gene-expression profiles and enriched pathways related to cytokine-cytokine receptor interaction, NF-κB, TNF, IL-17, Toll-like receptor, and NOD-like receptor signaling. The construct was immunogenic in mice; the exploratory MYY-containing formulation showed numerically higher immune-response readouts in this experimental setting, although these differences were not statistically significant. This adjuvant-associated observation requires further validation, and protection against MAP infection still needs to be tested in challenge experiments and target animals. Full article

Phycobiliproteins from the red alga Porphyra umbilicalis are rich precursors of bioactive peptides, but targeted identification of high-potency sequences remains challenging. To address this, we combined an in silico screening pipeline with experimental validation to discover novel antioxidant peptides. Virtual enzymatic hydrolysis generated 908 candidate sequences, which were filtered down to 12 peptides based on molecular weight, toxicity, hydrophilicity, and bioactivity predictions. Molecular docking and molecular dynamics simulations further identified four candidate peptides (FK, FKP, FRS, and RRF) that were computationally predicted to bind stably within the KEAP1 Kelch domain. Chemical assays confirmed that all four synthesized peptides exhibited dose-dependent DPPH and ABTS radical-scavenging activities, along with Fe²⁺-chelating capacity and ferric reducing antioxidant power. In H₂O₂-stimulated Caco-2 cells, peptide pretreatment protected against cellular oxidative stress by restoring endogenous antioxidant enzyme activities, increasing total antioxidant capacity, and reducing lactate dehydrogenase leakage. Among these, RRF displayed the strongest protective efficacy. Overall, this study presents a practical workflow for mining marine-derived peptides and highlights four candidate peptides whose potential as functional food ingredients warrants further validation in actual P. umbilicalis hydrolysates. Full article

Inflammatory disorders remain an important therapeutic challenge, highlighting the need to identify novel small molecules capable of modulating inflammatory signaling. In this study, an in-house library of aromatic and heterocyclic drug-like compounds was screened for inhibition of lipopolysaccharide (LPS)-induced nuclear factor kappa B (NF-κB) activation using THP-1 dual-reporter cells. Eight compounds markedly suppressed NF-κB activity, and five structurally related 4-phenylimidazole-based benzenesulfonamide derivatives (1232, 1239, 1241, 1242, and 1245) were prioritized based on their cytotoxicity profiles. Functional activity was subsequently evaluated in LPS-stimulated RAW 264.7 macrophages. The selected compounds reduced nitrite production, attenuated LPS-induced nuclear localization of NF-κB, and differentially affected IL-6, KC/CXCL1, and TNF-α production. The naphthyl-substituted derivative 1239 showed prominent inhibition of nitrite, IL-6, and KC/CXCL1 production, whereas the ethylthio derivative 1232 preferentially reduced TNF-α. Molecular docking against a panel of inflammation-associated proteins revealed favorable predicted interactions of compound 1239 with IκB kinase(IKK) and p38 mitogen-activated protein kinase (p38 MAPK), with docking scores of −10.3 kcal/mol for both proteins and binding poses overlapping the corresponding crystallographic ligand-binding regions. Collectively, these findings identify 4-phenylimidazole-based benzenesulfonamides as a promising chemical series with anti-inflammatory activity and suggest IKK and p38 MAPK as potential components of the affected signaling network. Further structure–activity and mechanistic studies are required to define the molecular targets and optimize the activity of this compound class. Full article

Herbal medicines (HMs) are chemically complex multi-component systems, yet analytical strategies that systematically connect chemical composition, biological activity, target prioritization, and quantitative analysis remain limited. Here, we established an integrated analytical workflow combining ultra-high-performance liquid chromatography–quadrupole time-of-flight tandem mass spectrometry (UPLC-QTOF-MS/MS), bioactivity-guided fractionation, network pharmacology, molecular docking, surface plasmon resonance (SPR), and quantitative ¹H NMR (qHNMR) to investigate Fu’an Granules. UPLC-QTOF-MS/MS provided a broad constituent-annotation profile of the formulation. Bioactivity-guided fractionation using an LPS-stimulated RAW 264.7 macrophage model showed that fractions FA3 and FA4 exhibited relatively stronger inhibition of nitric oxide (NO) production at the tested screening concentration. Corilagin, rutin, and isochlorogenic acid A were subsequently selected as representative activity-associated constituents according to their distribution in the chromatographic fractions and showed reduced NO production without detectable cytotoxicity under the tested conditions. Network pharmacology, protein–protein interaction analysis, pathway enrichment, and molecular docking prioritized EGFR and PTGS2 as candidate binding targets. SPR further demonstrated concentration-dependent interactions between the three representative constituents and both proteins, providing experimental support for ligand-target binding. An exploratory internal-standard-based qHNMR analysis estimated corilagin content at 5.7% (w/w) of the analyzed sugar-depleted extract. Collectively, this study presents an integrated workflow linking chemical profiling, activity-guided constituent prioritization, computational target analysis, biophysical binding assessment, and complementary quantitative analysis. The results support corilagin as a representative activity-associated quantitative constituent for Fu’an Granules while providing a general analytical framework for activity-oriented quality marker evaluation in complex herbal medicines. Full article

Characterised by progressive alveolar bone loss, periodontitis arises from immune dysregulation and is classified as a chronic inflammatory condition. Zerumbone (Zer) has anti-inflammatory properties; however, its mechanism within the periodontal context remains unclear. We investigated the therapeutic effects and potential molecular targets of Zer by integrating in vivo efficacy assessment (using male C57BL/6 mice), network pharmacology, molecular docking, and molecular dynamics simulations, together with analysis of public single-cell transcriptomic data, with mechanistic validation using RAW 264.7 macrophages stimulated with LPS. In vivo, Zer attenuated bone resorption, preserved collagen integrity, reduced the number of TRAP-positive osteoclasts, and mitigated inflammation without hepatorenal toxicity. Network pharmacology predicted NFKB1, MAPK14, and AKT1 as potential core targets, while IL-17 pathway enrichment suggested the involvement of IL17A; single-cell data localised NFKB1/MAPK14/AKT1 to macrophages and showed that IL17A is predominantly expressed in T cells. In vitro, Zer dose-dependently suppressed LPS-driven phosphorylation of NF-κB p65, p38 MAPK and AKT1, correlating with reduced pro-inflammatory cytokine release and an M1-to-M2 phenotypic shift. In vivo, Zer also reduced gingival IL-17A expression. However, as IL-17A is primarily produced by T cells and our in vitro experiments were conducted exclusively in macrophages, this observation remains correlative and does not establish a direct T-cell-mediated effect. Collectively, these findings suggest that Zer ameliorates periodontitis partly through suppression of macrophage NF-κB and p38 MAPK signalling, with reduced AKT1 phosphorylation observed alongside these effects; however, whether AKT1 causally contributes to these effects remains to be determined. In addition, the potential impact on T-cell-associated IL-17A responses warrants further investigation. Full article

Background/Objectives: Silver has long been used in medicine for its diverse effects. Coordination of this metal ion with bioactive ligands such as thiosalicylic acid derivatives allows the synthesis of potentially promising antimicrobial and antitumor agents. Methods: The in vitro antimicrobial activity of silver(I) complexes (C1–C5) and their corresponding ligands (L1–L5) was determined using a resazurin-based microdilution method, while the inhibition of preformed biofilms was assessed against S. aureus, P. mirabilis, and P. aeruginosa. To explore the binding modes and affinity of the investigated silver(I) complexes, a molecular docking study was performed against the LasR transcriptional regulator, while in silico ADMET screening was conducted using the pkCSM online platform. Finally, the evaluation of in vitro immunomodulatory effects of the silver(I) complexes was assessed by measuring cytokine concentrations and performing flow cytometry. Results: In vitro and in silico evaluations confirm that the C2 complex possesses an optimal structural balance of size and hydrophobicity, enabling superior antimicrobial and antibiofilm activity against P. aeruginosa. Regarding the immune response, treatment with the C3 complex decreased the concentrations of the proinflammatory cytokines IL-1β, TNF-α, IL-6, and IL-12, while increasing the concentration of the anti-inflammatory cytokine IL-10 after LPS stimulation. Conclusions: Our findings demonstrate that C2 primarily targets bacterial pathogens, while C3 exerts potent immunomodulatory and anti-inflammatory effects that differ between innate and acquired, naïve and primed immune system cells. Therefore, investigating the combined or sequential application of C2 and C3 may provide a promising strategy for the treatment of chronic biofilm-associated infections. Full article

Calycosin (CAL) is an isoflavone monomer derived from Astragalus membranaceus that possesses potent anti-inflammatory and antioxidative pharmacological activities. However, the specific role of CAL in lipopolysaccharide (LPS)-induced inflammatory injury in cardiomyocytes remains to be elucidated. In this study, we established an in vitro inflammatory injury model using LPS-stimulated H9c2 cardiomyocytes to evaluate the cardioprotective effects of CAL. We assessed cell viability, live/dead cell staining, oxidative stress markers, inflammatory cytokine profiles, and cardiac injury biomarkers to characterize the protective efficacy of CAL. Quantitative proteomics combined with bioinformatics analysis was employed to explore the underlying mechanisms. Molecular docking and molecular dynamics simulations were performed to analyze the predicted binding affinity and complex stability between CAL and the target proteins PPARγ and NF-κB. PPARγ and NF-κB mRNA and protein levels were measured by qPCR and Western blotting, and rescue experiments with the PPARγ-specific inhibitor GW9662 were performed to validate target dependency. Our results demonstrated that CAL exerted significant protective effects against LPS-induced cardiomyocyte injury, effectively enhancing cell viability, reducing cell death, alleviating intracellular ROS accumulation, modulating oxidative stress-related parameters, suppressing excessive inflammatory responses, and decreasing the release of cardiac injury biomarkers. Proteomic analysis revealed that differentially expressed proteins were significantly enriched in the PPAR signaling pathway. Molecular simulations predicted that CAL binds stably to both PPARγ and NF-κB. PCR and Western blotting results showed that CAL upregulated PPARγ and suppressed NF-κB-mediated pro-inflammatory gene expression at both the mRNA and protein levels. Notably, pharmacological blockade of PPARγ with GW9662 partially abolished the cardioprotective effects of CAL. Collectively, these findings indicate that CAL ameliorates LPS-induced inflammatory injury in H9c2 cardiomyocytes through a protective mechanism that depends on activation of PPARγ and subsequent inhibition of the NF-κB signaling pathway. Full article

Previous studies have reported that the combination of Astragali Radix and Angelicae Sinensis Radix (AR-ASR) can ameliorate atherosclerosis (AS) through anti-inflammatory effects; however, the exact mechanistic underpinnings still await further clarification. The present study investigates the anti-atherosclerotic action of AR-ASR from the perspective of PANoptosis (encompassing apoptosis, pyroptosis, and necroptosis). To ascertain the chief active ingredients of the AR plus ASR mixture, UPLC-MS/MS detection was utilized. To predict potential therapeutic targets, a multi-faceted bioinformatics strategy was applied, integrating transcriptomics, single-cell sequencing, network pharmacology, molecular docking, and molecular dynamics simulations. In vivo, male ApoE-deficient mice were allocated at random into the model cohort, AR-ASR low-, medium-, and high-dose groups (AR-ASR L, AR-ASR M, and AR-ASR H, respectively), atorvastatin group (ATV), and control group (age-matched wild-type C57BL/6J mice). Atherosclerosis was induced by 12 weeks of high-fat diet feeding, followed by 4 weeks of oral gavage treatment. Assessment parameters included aortic intimal hyperplasia thickness, serum lipid levels, vascular inflammatory factor expression, endothelial function, and the gene and protein expression levels of PANoptosis- and p38MAPK/p53 signaling pathway-related molecules. For in vitro experiments, POVPC-stimulated AVECs were used as an oxidative damage model and then treated with AR-ASR-containing serum. Cell proliferation activity, cell injury rate, and the expression levels of PANoptosis markers and p38MAPK/p53 signaling pathway-related molecules were measured. Furthermore, in the presence of a p38MAPK agonist, the protective effect of AR-ASR on AVECs and its underlying mechanism were investigated. Bioinformatics analysis suggested that AR-ASR modulates PANoptosis and interferes with AS via the p38MAPK/p53 signaling axis. In vivo, AR-ASR treatment reduced serum lipid levels, attenuated aortic intimal hyperplasia, decreased plasma inflammatory cytokine levels and vascular inflammatory factor expression, improved endothelial function, and suppressed both vascular PANoptosis and p38MAPK/p53 pathway activation. In vitro, POVPC induced oxidative damage and PANoptosis in AVECs. AR-ASR-containing serum promoted AVEC viability, reduced LDH release, inhibited the expression of PANoptosis-related markers, and downregulated p38MAPK/p53 signaling pathway protein expression. Furthermore, in the presence of a p38MAPK agonist, the inhibitory effects of AR-ASR on cellular PANoptosis and p38MAPK/p53 pathway activation were significantly attenuated. The AR-ASR compatibility suppresses p38MAPK/p53 pathway activation, thereby inhibiting vascular endothelial PANoptosis-driven inflammatory responses and ameliorating atherosclerotic vascular pathology. Full article

Food spoilage and foodborne microorganisms remain major challenges for food safety and preservation, stimulating the search for alternative antimicrobial agents compatible with minimally processed and clean-label products. In this study, six short synthetic peptides containing arginine, or both lysine and arginine residues, were evaluated as potential antimicrobial agents against Staphylococcus aureus, Salmonella enterica, and Candida albicans. Minimum inhibitory concentrations were determined by broth microdilution, and molecular docking was performed against C. albicans lanosterol 14α-demethylase to provide molecular-level insights. None of the peptides inhibited S. enterica within the tested concentration range, indicating limited activity against the Gram-negative bacterium. Peptides 1, 3, 5, and 6 inhibited S. aureus only at relatively high concentrations, with MIC values between 1598.7 and 1782.0 µmol L⁻¹, without confirmed bactericidal activity. In contrast, peptide 4, FEFKFEFKGGGRGDS, was the only sequence active against C. albicans, showing an MIC of 19.93 µmol L⁻¹. However, no minimum inhibitory concentration (MIC) was obtained within the tested concentration range, indicating that the observed antifungal effect should be interpreted as inhibitory rather than confirmed fungicidal activity. Docking analysis was consistent with this experimental trend, as peptide 4 exhibited the most favorable predicted binding energy, −8.4 kcal mol⁻¹, and established predicted contacts with catalytic site of ERG11, including Lys143, Arg381, His468, Arg469, and Cys470. These findings indicate that antimicrobial activity was strongly sequence- and microorganism-dependent. Peptide 4 represents a promising antifungal lead sequence for further optimization and validation in food-relevant systems. Full article

GPR119 is a promising target for the treatment of type 2 diabetes mellitus. Stimulating this receptor leads to the induction of glucose-dependent insulinotropic peptide and glucagon-like peptide 1, which improves systemic glucose homeostasis. Encouraged by our previous research, we replaced 5-nitropyrimidine with 1,3,5-triazine to reduce hepatotoxicity. A new series of compounds with a 1,3,5-triazine scaffold was synthesized and evaluated for their agonistic activities against human GPR119. Thirty-three compounds were synthesized and evaluated for their potential GPR119 agonist activities. Analogs with a 4-cyano and a 4-methylsulfonyl group in the aryl ring exhibited more potent activation than those analogs with a 4-trifluoromethyl group. Compound 3c had the most favorable EC₅₀ value (hEC₅₀ = 2.39 µM) and significantly induced glucagon-like peptide-1 secretion. Furthermore, docking simulations showed that compound 3c stably binds to the active site of GPR119. Full article

Transient receptor potential vanilloid channel 3 (TRPV3) is a keratinocyte-expressed ion channel that regulates calcium signaling and thymic stromal lymphopoietin (TSLP) production, contributing to pruritic inflammatory skin diseases. Although avicularin (AVC), a flavonoid from Polygonum aviculare L., and senegenin (SNG), the bioactive aglycone of Polygala tenuifolia Willd., possess anti-inflammatory properties, their effects on TRPV3-associated signaling remain unknown. A natural product library was screened for nitric oxide (NO) production in carvacrol-stimulated human keratinocytes. Following primary screening and secondary evaluation of TRPV3 and TSLP expression, AVC and SNG were selected for further study and subsequently evaluated by molecular docking, calcium imaging, quantitative real-time PCR, immunocytochemistry, and pathway inhibition assays. AVC and SNG showed favorable predicted docking poses at both TRPV3 docking sites and significantly attenuated carvacrol-induced calcium-dependent fluorescence responses in TRPV3-expressing cells. Both compounds reduced TRPV3 and TSLP expression at the mRNA and protein levels and attenuated the nuclear translocation of phospho-Nuclear Factor of Activated T Cells 2 (NFATC2) and phospho-p50. Co-treatment with NFAT or Nuclear Factor Kappa B (NF-κB) inhibitors produced no additional suppression, consistent with convergence of AVC/SNG-associated effects with NFAT/NF-κB signaling. Collectively, these findings support further investigation of AVC and SNG as candidate modulators of TRPV3-associated inflammatory signaling in keratinocytes. Full article

Persistent inflammation driven by excessive AKR1B1 activation contributes to the progression of multiple chronic inflammatory diseases. Natural chlorogenic acid (CGA) exhibits anti-inflammatory activity but suffers from poor membrane permeability, while its laurate ester derivative (CGL) shows clear dose-dependent cytotoxicity in macrophages. Here, we synthesized a novel chlorogenic acid amide derivative (CGAA) by selective amidation of the aliphatic carboxyl group on the quinic acid moiety of CGA, fully preserving the chiral polyphenol scaffold. Structural identity was confirmed by HSQC, HMBC, FTIR and HR-ESI-MS. CCK-8 assays demonstrated that CGAA maintained > 95% viability of RAW264.7 cells at 100 μM, displaying markedly superior cellular safety relative to CGL. In LPS-stimulated RAW264.7 macrophages, CGAA dose-dependently suppressed NO, TNF-α and IL-6 production and exhibited stronger antioxidant activity than both CGA and CGL. Temperature-gradient CETSA, DARTS and enzymatic assays established direct binding and potent inhibition of AKR1B1 by CGAA (IC₅₀ = 7.85 μM). Molecular docking revealed that the newly introduced amide forms an additional hydrogen bond with ILE-260, enhancing ligand–target affinity. This carboxyl-amidation strategy simultaneously improves the cellular safety and AKR1B1 inhibitory potency of CGA, providing a rational approach for developing low-toxicity polyphenol-based anti-inflammatory leads. Full article

Phytochemicals modulate lipid metabolism, but it remains unclear whether phytochemical-rich cactus fruits have a similar effect. Accordingly, the effects of Etcho (Pachycereus pecten-aboriginum) fruit juice on lipid accumulation and on the expression of CD36 and ADIPOR1 were investigated, and the potential interactions between its major betalains and the CD36 and ADIPOR1 receptors were predicted. Phytochemical content was determined by spectrophotometry. Human adipocytes were stimulated with a 2:1 (molar) oleic/palmitic acid mixture and then treated with Etcho fruit juice/its hydroethanolic extract/betanin. Lipid accumulation was measured by Oil Red O staining, and CD36 and ADIPOR1 expression were quantified by qPCR. Molecular docking was performed to assess the binding affinities and molecular interactions of betanin, indicaxanthin, and phyllocactin with CD36 and ADIPOR1. All treatments reduced long-chain fatty acid–induced lipid accumulation (p < 0.05). The juice decreased CD36 mRNA and increased ADIPOR1 mRNA levels (p < 0.05), and molecular docking revealed that its betalains bind strongly to the CD36 extracellular domain and the ADIPOR1 orthosteric pocket triad. Etcho fruit juice is a source of phenolics and betalains. It reduces lipid accumulation in human adipocytes and modulates the expression of CD36 and ADIPOR1. The juice’s major betalains may target CD36 and ADIPOR1. Full article

The hepatocyte growth factor (HGF)/c-Met signaling pathway plays a central role in cellular proliferation, survival, migration, invasion, angiogenesis, and therapeutic resistance. Aberrant c-Met activation, driven by gene amplification, overexpression, activating mutations, exon 14-skipping alterations, or ligand-dependent stimulation, drives the development and progression of many solid tumors, positioning c-Met as a key target for anticancer drug development. The clinical effectiveness of c-Met-targeted treatments such as crizotinib, capmatinib, tepotinib, savolitinib, and cabozantinib has confirmed c-Met as a viable oncogenic driver for therapy, leading to the development of various next-generation inhibitors with different structures. This review provides a comprehensive perspective on small-molecule c-Met inhibitors from the perspectives of medicinal chemistry and structure-based drug design, encompassing approved drugs, investigational agents, natural-product-inspired leads, and emerging multitarget anticancer therapeutics. Particular emphasis is given to the principles of molecular recognition that govern c-Met inhibition. This includes the structure of the kinase domain, interactions at the ATP-binding site, recognition of the hinge region, and the different binding modes of Type I, Type II, and allosteric inhibitors. The design, synthesis, biological evaluation, and structure–activity relationships of diverse heterocyclic scaffolds that have shaped c-Met inhibitor discovery are critically analyzed. Key medicinal chemistry strategies, including scaffold hopping, bioisosteric replacement, conformational optimization, molecular hybridization, and multitarget pharmacophore integration, are discussed in the context of potency, selectivity, resistance management, and drug-like properties. Particular attention is given to the integration of structural biology, molecular docking, binding-mode analysis, and structure-guided optimization approaches that have enabled the development of potent c-Met-directed inhibitors. In addition, recent advances in dual- and multitarget agents that simultaneously modulate c-Met and complementary therapeutic targets, including VEGFR-2, EGFR, AXL, MER, PARP1, CDK2, and tubulin, are highlighted as promising strategies for overcoming pathway redundancy and acquired resistance. This review summarizes contemporary structure-based and medicinal chemistry principles underlying c-Met inhibitor discovery, critically evaluates the relationship between biochemical potency and therapeutic efficacy, and provides a framework for the rational design of next-generation c-Met-targeted and multitarget anticancer agents. Full article

The increasing prevalence of biofilm-associated infections caused by Cutibacterium acnes has stimulated interest in food-derived natural products as alternative sources of anti-biofilm agents. This study investigated the effects of processing and extraction conditions on the phytochemical composition, antibacterial activity, and anti-biofilm properties of Dioscorea hispida Dennst. Reflux extraction of dried yam with 80% ethanol produced the crude extracts with the highest yields (1.39–1.80%), whereas fresh yam yielded 0.51–0.97% extract. Using Gas–liquid chromatography–mass spectrometry (GLC-MS) analysis, linoleic acid ethyl ester, n-hexadecanoic acid, 9,12-octadecadienoic acid (Z,Z)-, and stigmasterol were identified as the major constituents. Among the tested extracts, DH-W-F-H (D. hispida-water washing-fresh-hexane) and DH-W-F-E (D. hispida-water washing-fresh-ethanol) were extracted from fresh yam using hexane and ethanol, respectively, while DH-W-D-E (D. hispida-water washing-dry-ethanol) was isolated from dried yam using ethanol and exhibited the strongest antibacterial activity, with minimum inhibitory concentrations (MIC) ranging from 64 to 2048 µg/mL. These extracts demonstrated pronounced concentration-dependent inhibition of biofilm formation by Staphylococcus epidermidis, Staphylococcus aureus, and Cutibacterium acnes. The strongest anti-biofilm activity was observed against C. acnes, with biofilm formation nearly eliminated at MIC concentrations. Moreover, all three extracts significantly reduced established C. acnes biofilms, with DH-W-F-H exhibiting greater eradication efficacy than vancomycin under the tested conditions. To elucidate the underlying mechanism, major fatty acid derivatives were evaluated against C. acnes lipase (_CALipase), a virulence factor associated with biofilm development, using molecular docking, molecular dynamics simulations, and the Molecular Mechanics-Generalized Born Surface Area (MM-GBSA) binding free-energy calculations. The compounds exhibited favorable interactions with _CALipase, with linoleic acid ethyl ester (FA2) showing the strongest binding affinity, stable protein–ligand interactions throughout a 200 ns simulation, and the most favorable binding free energy. Collectively, the biological and computational findings suggest that fatty acid-rich extracts from processed D. hispida suppress biofilm formation through an antivirulence mechanism involving _CALipase inhibition. These results highlight the potential of D. hispida as a source of metabolites for the development of functional food ingredients and value-added cosmetic and dermatological applications. Full article