Construction and biological evaluation of novel carbazole-5-phenyl-1,3,4-oxadiazole derivatives as multi-target hypoglycemic agents.

Discovery of the first dual PD-L1/JAK inhibitor with enhanced in vivo antitumor immunity.

Fifty-one thymol-based chalcone derivatives were designed and evaluated for their antifungal activities against eight plant pathogenic fungi in vitro. Compound Q23 exhibited significant inhibitory activity against Valsa mali (V. mali), with an EC50 value as low as 1.52 μg/mL, which was markedly superior to that of the lead compound thymol (EC50 = 13.7 μg/mL) and reference fungicide azoxystrobin (Az, EC50 = 21.9 μg/mL). In vivo experiments further confirmed that compound Q23 effectively controlled apple canker caused by V. mali. Preliminary mechanism studies revealed that compound Q23 induced the accumulation of reactive oxygen species in fungal cells, leading to membrane lipid peroxidation, disruption of cell membrane integrity and physiological functions, and ultimately cell death. These findings suggest that compound Q23 is a promising candidate fungicide for the control of apple canker.

The topoisomerase 1 (TOP1) enzymatic inhibition and antiproliferative activity of phosphorated indenoquinoline derivatives were investigated. First, the preparation of new hybrid quinoline and tetrahydroquinoline structures with a phosphine oxide group was performed by a two-step Povarov type [4 + 2]-cycloaddition reaction between the corresponding phosphorated aldimines with indene in the presence of BF3·Et2O, affording corresponding 1,2,3,4-tetrahydroindeno[2,1-c]quinolinylphosphine oxides 9, 7H-indeno[2,1-c]quinolinylphosphine oxides 10 and 7-oxoindeno[2,1-c]quinolinylphosphine oxides 11 with good yields. The synthesized derivatives were evaluated as TOP1 inhibitors, showing that some derivatives (9f, 9g, 9l, and 11m) show better or similar activity to the reference compound (CPT) at 1 min. The synthesized derivatives were screened for their antiproliferative activity in different cancerous cell lines, and all of them present a higher selective cytotoxicity in the human lung adenocarcinoma cell line (A549), than in the others. In contrast, almost none of the synthesized phosphorated compounds exhibited antiproliferative activity toward nonmalignant lung fibroblasts MCR5. These results suggest that phosphine oxide-substituted quinoline derivatives have important properties as TOP1 inhibitors and show an interesting cytotoxicity against six different cancerous cell lines.

Natural genetic variation impacts complement inhibitory activity of PFam54 orthologs of Asian Borrelia bavariensis.

Lysine-specific demethylase 1 (LSD1) is a crucial target due to its essential role in cancers. CC-90011, the reversible and highly selective LSD1 inhibitor at the clinical stage, has demonstrated significant therapeutic potential in acute myeloid leukemia and solid tumors. Herein, we present a novel synthetic route to LSD1-targeting compounds that enables late-stage diversification; the key intermediate LYY-23 was used to synthesize CC-90011 and MS9117. In addition, compounds LYY-25 ∼ LYY-29 were synthesized and evaluated for LSD1 inhibition, all of which showed inhibitory activity, with LYY-28 emerging as the most potent compound (IC50 = 3.59 nM).

Butyrate-producing bacteria (BPB) are widely used as additives in aquaculture because of the beneficial effects of the butyrate they produce. However, most available BPB currently are of terrestrial origin, which are quite different from aquatic animals, resulting in profoundly unstable probiotic performance. This study isolated a novel, indigenous BPB, Clostridium moniliforme G18, from the gut of mud crabs, Scylla paramamosain. The strain demonstrated robust butyric acid production, tolerance to gastrointestinal conditions (pH 4-9, 0-3% NaCl, 0.4-1.0% bile salts), antibiotic sensitivity, pathogen inhibition, and non-hemolytic activity. Dietary supplementation with G18 alone or in combination with resistant starch and galactooligosaccharides (GR18) significantly enhanced mud crab body weight (P < 0.05). The supplementation improved intestinal health by enhancing histomorphology and upregulating tight junction genes (ZO-1 and Claudin). Furthermore, it boosted antioxidant capacity (elevated SOD and CAT; reduced MDA), modulated immune responses (increased AKP and ACP; reduced ALT and AST, P < 0.05), and increased intestinal butyrate levels. Analysis of the gut microbiota of mud crabs fed G18 alone or in combination with prebiotics revealed an increase in alpha diversity and beneficial composition changes (including reduced Vibrionaceae and elevated Proteocatella, P < 0.05). Following Vibrio parahaemolyticus challenge, mud crabs fed G18 and GR18 showed significantly higher survival rate (66.7% and 73.3%, respectively, P < 0.05) versus 53.3% in the control group. These diets mitigated intestinal damage, restored immune and antioxidant parameters, and enhanced the expression of barrier function genes. These results establish indigenous C. moniliforme G18 as a highly promising probiotic candidate for enhancing growth and health in mud crabs.

This study investigated the anti-inflammatory and skin barrier-improving activities of Fructobacillus fructosus strain NSH-1-derived exosomes in LPS-stimulated RAW 264.7 cells and HaCaT cells. Nanoparticle tracking and transmission electron microscopy analyses confirmed the concentration, purity, and morphology of NSH-1 exosomes. The exosomes showed no cytotoxicity at concentrations of 1.5 × 107, 3.0 × 107, and 6.0 × 107 particles/ml, but demonstrated concentration-dependent inhibitory activity against nitric oxide production. Furthermore, NSH-1 exosomes significantly inhibited the production of pro-inflammatory mediators, including TNF-α, IL-6, IL-1β, and PGE2. In HaCaT cells, treatment with NSH-1 exosomes enhanced skin barrier function by increasing the production of hyaluronan and collagen in a concentration-dependent manner and promoting wound-healing activity. Western blot analysis revealed that treatment with NSH-1 exosomes reduced the expression levels of inflammatory proteins iNOS and COX-2, while increasing the expression of skin barrier-related proteins such as filaggrin, involucrin, and loricrin. Overall, these findings indicate that NSH-1 exosomes exert strong anti-inflammatory activity by suppressing the expression of the pro-inflammatory cytokines iNOS and COX- 2, while simultaneously enhancing skin barrier integrity by upregulating filaggrin, involucrin, and loricrin. Thus, exosomes derived from F. fructosus NSH-1, isolated from Campsis grandiflora flowers, demonstrate promise as natural bioactive agents for anti-inflammatory and skin barrier-improving applications.

Experimental evaluation of an integrated focused ultrasound and electroencephalography approach for developing activation-informed neuroimaging.

Thioamides constitute an important class of pharmaceutically active natural products, yet their discovery and development are limited. A targeted metabologenomic method is developed to logically and efficiently discover thioamide compounds in bacteria. To this end, two strains were identified to possess genetic capacity for biosynthesizing thioamides from the bacterial genomic DNA library (1,192 strains) using the polymerase chain reaction to target the TfuA-encoding gene, a genomic hallmark of thioamide biosynthesis. Mass spectrometric isotopic patterns of sulfur-bearing compounds serve as metabolomic hallmarks to detect thioamide production from the extracts of the selected strains without chromatography. Applying this metabologenomic targeting approach, two new thioamides, thiogochangamides A and B, belonging to the thioviridamide family whose stereochemistry has remained unresolved for two decades, were discovered in Streptomyces sp. Their absolute configurations were fully assigned through chemical derivatizations, including the advanced Marfey method, Mosher method, partial hydrolysis, synthesis of an unusual amino acid, and desulfurization, combined with computational methods. Thiogochangamide B exhibits potent inhibitory activity against gemcitabine-resistant pancreatic cancer cells both in vitro and in vivo. Mechanistically, thiogochangamide B effectively downregulates Wnt/β-catenin signaling, thereby suppressing the metastatic potential of drug-resistant cancer cells. This study provides a new therapeutic strategy for overcoming recalcitrant drug-resistant pancreatic cancer.

Breast cancer is the most often diagnosed malignancy in women, exceeding lung cancer with almost 2.3 million new cases per year. Triple-negative breast cancer (TNBC), comprising 10%-15% of cases, is distinguished by its aggressive development, high invasiveness, and restricted treatment alternatives. Patients with malignancies often exhibit impaired immune function, predisposing them to opportunistic infections, particularly those caused byS. aureus (S. aureus). In this study, the development and characterization of a multifunctional liposomal nanosystem co-delivering betanin and IR806 dye (Bet-IR806-LPs) for the dual management of TNBC and bacterial infections was reported. The liposomes were formulated using soybean phosphatidylcholine (Soya PC), which enhanced drug encapsulation, stability, and biocompatibility. Bet-IR806-LPs exhibited strong photothermal (PTT) conversion efficiency under near-infrared (NIR) irradiation, enabling effective PTT and photodynamic therapy (PDT). Antibacterial assessment againstS. aureusrevealed potent inhibitory activity, with an IC50of 5 μg ml-1. Biocompatibility was confirmed in fibroblast cell models, while zebrafish assays provided further insights into nanomaterial safety through analyses of reactive oxygen species and apoptosis. Cytotoxicity studies in 4T1 TNBC cells demonstrated significant anti-proliferative and anti-migratory effects, with enhanced tumor cell ablation upon NIR irradiation mediated by combined PTT-PDT mechanisms. Collectively, Bet-IR806-LPs demonstrate promising dual functionality, integrating antibacterial and anticancer activities within a single nanosystem. These findings suggest their potential as a versatile therapeutic platform with clinical relevance for improving treatment outcomes in TNBC while simultaneously addressing bacterial co-infections.

A novel series of antioxidant-conjugated mutual prodrugs of ibuprofen (RJ-02-01 to RJ-02-10) was designed and synthesized to address the gastrointestinal (GI) toxicity. Ibuprofen was covalently linked to substituted aniline-based antioxidant moieties through an amide bond. In-vitro hydrolysis results demonstrated the immense stability of the synthesized ibuprofen prodrugs in simulated gastric fluid (pH 1.2) with minimal drug release, indicating reduced potential for gastric irritation. Whereas enhanced hydrolysis was observed in simulated intestinal fluid (pH 7.4). Molecular docking revealed strong binding affinities for selected derivatives, with RJ-02-07 showing the most favorable interaction profile compared with ibuprofen. Biological evaluation identified RJ-02-07 as the lead compound, exhibiting significantly stronger COX-1 and COX-2 inhibitory activity than the parent drug. Cellular assays further demonstrated reduced reactive oxygen species levels, increased superoxide dismutase activity, and downregulation of COX-1/COX-2 protein expression, indicating improved antioxidant and anti-inflammatory potential. Cytotoxicity assessment showed lower toxicity relative to ibuprofen, suggesting a favorable safety profile. These findings suggest that this mutual prodrug strategy may reduce gastric exposure to free ibuprofen while enhancing the therapeutic efficacy; however, further in-vivo pharmacokinetic and gastro-protective studies will be required to confirm these potential benefits.

This study identified novel DPP-IV inhibitory oligopeptides from bovine casein hydrolysates for hyperglycemia regulation. Initial in vitro screenings showed that low-molecular-weight fractions (<3 kDa) exhibited superior DPP-IV inhibitory activity. From these fractions, high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) integrated with in silico, in vitro, and in situ validations identified potent oligopeptides, notably FPQYLQYL, which outperformed FVAPF and DAYPSGAWY. Kinetic analysis confirmed a competitive inhibition mode targeting the catalytic pocket. Innovatively, integrating microscale thermophoresis (MST), fluorescence quenching, molecular dynamics simulations, and atomic force microscopy (AFM) proved that the inhibitory potency was driven by high binding affinity and oligopeptide-enzyme aggregation. Crucially, structure-activity relationship analysis identified a critical substrate-mimicking N-terminal Phe-Pro motif, where the proline-driven insertion is stabilized by specific hydrophobic and aromatic interactions within the pocket. Collectively, our findings provided insights into the design and discovery of bioactive oligopeptides with hypoglycemic potential and offered a promising natural alternative to synthetic DPP-IV inhibitors.

Ice-binding proteins (IBPs) are crucial for the survival of cold-adapted organisms, as they regulate ice crystal formation and growth. To understand their molecular mode of action, fluorescence microscopy of IBPs bound to ice-crystal surfaces has been shown to be very helpful, although it is unknown whether the (fluorescent) tags typically used in these studies affect the activities of the IBPs. Here, we evaluate the impact of mEos3.2, SNAP-tag, and HaloTag on the ice-recrystallization inhibition (IRI) activity of IBPs. We find that most tags, in most orientations, do not affect the IRI activity of IBPs. These tags are promising candidates for investigating the binding mechanisms of IBPs in their native form with fluorescence microscopy. A surprising exception is the N-terminal attachment of HaloTag to QAE, an isoform of AFP type III: for this case, we find an order of magnitude higher IRI activity. Additionally, we show that HaloTag also has moderate IRI activity by itself and induces the formation of ice crystals with hexagonal prism morphology, suggesting binding affinity for the primary prism plane of ice. Our findings indicate that moderately IRI-active proteins may synergistically enhance the IRI activity of IBPs, when attached in the proper orientation.

The emergence of New Delhi metallo-β-lactamase-1 (NDM- 1) in Escherichia coli has become a significant public health threat due to its ability to hydrolyze a broad range of β-lactam antibiotics, including carbapenems. This study aimed to identify natural inhibitors of NDM-1 using a combined in silico and in vitro approach, with a focus on phytoconstituents from medicinal plants. Five medicinal plants were screened for antibacterial activity against an NDM-1- producing E. coli strain. Extracts of Curcuma longa L. prepared with acetone, ethanol, and water were evaluated using an agar well diffusion assay. A phytochemical database of C. longa was compiled, and 144 compounds were assessed for drug-likeness using SwissADME. Compounds meeting selection criteria were subjected to molecular docking against the NDM-1 protein. The top hit was further evaluated for antibacterial and synergistic activity with amoxicillin. Minimum Inhibitory Concentration (MIC) was determined, and in silico toxicity and pharmacokinetic profiles were predicted. C. longa acetone extract exhibited the highest antibacterial activity, with inhibition zones ranging from 8 to 16 mm. Ninety-six compounds passed drug-likeness filters; bisdesmethoxycurcumin showed the best binding energy (-6.5 kcal/mol) with NDM-1. In vitro, it demonstrated synergistic activity with amoxicillin, increasing inhibition zones from 10.33 mm to 19.67 mm. MIC was found to be 25-30 μg/ml. Toxicity predictions suggested acceptable safety and pharmacokinetic parameters. The study highlights the potential of bis-desmethoxycurcumin as an adjuvant to conventional antibiotics to overcome NDM-1-mediated resistance. Bis-desmethoxycurcumin from C. longa exhibits promising inhibitory activity against NDM-1-producing E. coli, supporting its potential use in adjuvant therapy to combat antibiotic resistance.

Background/Objectives: This study provides the first comprehensive evaluation of the antioxidant, antimicrobial, and enzyme inhibitory activities of Onosma alboroseum subsp. alboroseum var. alboroseum, including a novel nanoformulation-based comparative assessment of its most active extract. The study further aimed to investigate whether nanoparticles modulate the biological performance of the extract. Methods: Antioxidant activity was assessed using DPPH, FRAP, and CUPRAC assays, and total phenolic content was determined by the Folin–Ciocalteu method. Antimicrobial activity was evaluated using agar well diffusion and microdilution assays, while enzyme inhibitory activities were assessed through anticholinesterase and anti-urease assays. The most biologically active extract was subjected to LC–QTOF–MS-based tentative metabolite profiling and subsequently formulated into nanoparticles for comparative biological evaluation. Results: Among the extracts studied, the methanol extract had the highest total phenolic content and demonstrated superior antioxidant, antimicrobial, and enzyme inhibitor activities. LC–QTOF–MS profiling indicated a phenolic-rich composition, with rosmarinic acid as the predominant compound based on relative peak area. The methanol extract was encapsulated within alginate nanoparticles for subsequent comparative biological assessment. While the crude extract showed superior activity in antioxidant assays, nanoparticles enhanced cholinesterase and urease inhibition (28.03% and 12.11%, respectively) and improved antibacterial efficacy in microdilution assays (MIC range: 3.13–12.5 µg/mL), although no inhibition was observed in agar diffusion tests. Conclusions: These findings indicate the first time that the methanol extract of Onosma alboroseum subsp. alboroseum var. alboroseum represents a phenolic-rich source of bioactive constituents and a nanoparticle formulation that can modulate specific biological activities depending on the assay system, highlighting the relevance of formulation strategy in phytochemical-based pharmaceutical applications.

Zika virus (ZIKV) infection in adults is typically mild and self-limiting, but infection during pregnancy can cause fetal and neonatal microcephaly. No approved drugs or vaccines currently exist. ZIKV hijacks host lipid homeostasis to facilitate replication, and inhibitors of fatty acid synthesis decrease its production. However, cells obtain lipids from both de novo synthesis and uptake. Neurons, a primary target of ZIKV pathogenesis, have limited capacity for de novo lipid synthesis and rely more on lipid uptake. We previously demonstrated that lipid uptake inhibitors block influenza virus replication, yet their efficacy against ZIKV-particularly in neuronal cells-remains unknown. In the present study, we examined the anti-ZIKV and fatty acid uptake inhibitory activities of lipofermata, a fatty acid transporter protein isoform 2 (FATP2)‒specific inhibitor. Lipofermata effectively inhibited ZIKV production in immortalized hepatocyte (imHC) and SH-SY5Y (neuroblastoma) cell lines, with respective half-maximal inhibitory concentration (IC50) values of 1.41 and 1.11 μM with 50% cytotoxic concentration (CC50) of more than 20.0 μM. FATP2 mRNA and protein expression was significantly elevated in infected SH-SY5Y cells. ZIKV also significantly increased cellular phosphatidylcholine content. Lipofermata inhibited fatty acid uptake with an IC50 of 7.0 μM and significantly decreased phosphatidylcholine level. In addition, FATP2 knockdown by siRNA significantly decreased FATP2 mRNA and protein expression in SH-SY5Y and decreased ZIKV titer. These findings suggest that lipofermata disrupts ZIKV replication via fatty acid uptake inhibition. Furthermore, lipofermata at 2 µM markedly diminished caspase 3/7 and 8 activities, indicating that lipofermata mitigated ZIKV-induced extrinsic apoptosis in SH-SY5Y. Mitigating apoptosis may result in reduced neuropathogenesis. Consequently, FATP2 may serve as a promising target for anti-ZIKV therapies.IMPORTANCEZika virus can infect and cause severe damages to the brain. There is no effective antiviral drug for the treatment of Zika infection. Similar to many viruses, replication of Zika virus requires lipid. We show here that the virus could upregulate the fatty acid transporter FATP2 in neuroblastoma cells, which may increase the lipid supply in the infected cells, and that inhibiting the fatty acid transporter by lipofermata could suppress the viral replication and apoptosis. ZIKV production was also reduced by FATP2 knockdown. These suggest that FATP2 may be a promising target for the development of anti-Zika medication.

Species of the genus Stachys (Lamiaceae) are recognized for their ethnobotanical importance and chemical diversity. In this study, the essential oil (EOS) and solvent extracts of the endemic species Stachys sparsipilosa were investigated using integrated GC–MS and LC–ESI–QTOF/MS approaches. GC–MS analysis showed that identified constituents accounted for 94.62% of the total oil, with caryophyllene oxide, kauran-16-ol, and cubebol as major components. Targeted LC–MS analysis quantified prominent phenolic compounds, including chlorogenic acid, rutin, and hesperidin, while untargeted metabolomics tentatively annotated 168 metabolites belonging to phenolics, terpenoids, and other classes. Antioxidant capacity was evaluated using complementary in vitro assays, and enzyme inhibitory activities against α-amylase, α-glucosidase, tyrosinase, acetylcholinesterase, and butyrylcholinesterase were assessed in comparison with standard inhibitors. The extracts demonstrated measurable but generally moderate activities relative to the corresponding positive controls. The essential oil exhibited moderate, non-selective cytotoxic effects at relatively high concentrations, whereas solvent extracts showed limited activity within the tested range. Molecular docking analyses were performed as supportive tools to explore possible enzyme–ligand interactions. Overall, S. sparsipilosa displays a chemically diverse metabolite profile associated with composition-dependent bioactivities, providing a basis for further mechanistic and in vivo studies.

Objective Adipocyte Fatty Acid-Binding Protein 4 (FABP4) exerts a direct negative inotropic effect on cardiac muscle, but the underlying cellular mechanisms remain elusive. This study aimed to dissect the specific effects of FABP4 on the contractility and calcium (Ca 2+ ) homeostasis of isolated mouse ventricular myocytes and to characterize the functional role and critical residues of its N-terminal domain. Methods Contractility and intracellular Ca 2+ transients were simultaneously measured in isolated adult mouse ventricular myocytes using an IonOptix system following acute application of recombinant human FABP4 or its synthetic N-terminal peptide (FABP4 aa1-20 ). L-type Ca 2+ current was assessed via the whole-cell patch-clamp technique. Dose-response curves were analyzed using non-linear regression, and site-directed mutagenesis (E15K) was performed to evaluate the functional importance of a key amino acid residue. Results FABP4 inhibited myocyte contraction in a biphasic, dose-dependent manner, with a high-affinity (EC 50 = 0.010 pM) and a low-affinity (EC 50 = 0.120 nM) component. This inhibition was largely independent of Ca 2+ handling, as Ca 2+ transient amplitude was only weakly attenuated at higher concentrations (EC 50 = 0.412 nM), and L-type Ca 2+ current was unaffected. In stark contrast, the FABP4 aa1-20 peptide also inhibited contraction (EC 50 = 0.110 nM) but did so via a Ca 2+ -dependent pathway, robustly suppressing Ca 2+ transients. Mutation of glutamic acid at position 15 (E15K) significantly attenuated the peptide’s inhibitory activity. Conclusion Full-length FABP4 suppresses cardiomyocyte contractility primarily through a Ca 2+ -independent pathway, likely by reducing myofilament Ca 2+ sensitivity. Conversely, its isolated N-terminal domain operates via a distinct, Ca 2+ -dependent mechanism. These findings reveal a complex dual-pathway regulation of cardiac function by FABP4 and identify its N-terminal region as a potential therapeutic target for mitigating obesity-related cardiac dysfunction.

Acylphloroglucinol derivatives with α-glucosidase inhibitory activity from Eucalyptus grandis × urophylla fruits.