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Matrix metalloproteases (MMPs), which are activated during malignancy growth and metastasis, have been a focus of current anticancer research and development. The selective recognition and precise inhibition of active MMPs could limit cancer progression. Photodynamic therapy (PDT) is a well-established local treatment for solid tumors. MMPs are expressed primarily in the vicinity of the tumor, and PDT strongly influences this process. However, in rare cases, PDT can activate MMPs. An improved PDT outcome was observed with the action of an MMP inhibitor (MMPI), namely Prinomastat. Research on this topic remains limited, presenting a substantial opportunity for further investigations. Various small-molecule compounds have been designed to target MMPs as potential inhibitors, but clinical trials have not confirmed their efficacy in vitro or in vivo. Currently, novel MMPIs with complex chemical structures are being designed and have shown high efficacy in initial preclinical studies. This review aims to provide a critical overview of recent advances in the use of cancer-related MMPs as therapeutic targets, along with new innovative approaches to targeting them. The effects of PDT on cancer-related MMPs, along with the advantages of combining therapies that could enhance curative efficacy, are discussed. The novel inhibitory approaches that regulate cancer-related MMPs are summarized. Full article

Intranasal administration is a promising drug delivery route enabling precise and rapid central nervous system targeting. In our previous work, twelve hybrid colloidal dispersions were developed, consisting of synthetic poly(ethylene-oxide)-b-poly(ε-caprolactone) (PEO-b-PCL) block copolymers with an increasing proportion of the hydrophobic PCL segment, Tween 80 (Tw80) and β-cyclodextrin derivatives (βCD), either methyl-β-CD (MβCD) or hydroxy-propyl-β-CD (HPβCD) for IN delivery of ropinirole hydrochloride (RH). Colloidal dispersions were prepared at different weight ratios (system/RH equal to 10:1 and 10:5), characterized and evaluated in vitro. The aim of this study is to evaluate the ex vivo permeation through rabbit nasal mucosa and determine the pharmacokinetic parameters of RH, when administered intranasally as a colloidal dispersion, compared with oral and intranasal RH solutions in C57BL/6J mice. Ex vivo permeation studies showed that all formulations significantly enhanced RH permeation compared to the pure RH solution (0.5 mg/mL, pH 5.6). Among them, F4 [(PEO-b-PCL₁/Tw80/HPβCD)/RH 10:5] was selected for further investigation. Pharmacokinetic analysis showed that F4 significantly enhanced both systemic and brain exposure of RH, achieving higher serum AUC and C_max values, despite a 3-fold lower administered dose compared to the oral dose. It showed high systemic (F_rel(Serum) = 1815%) and brain (F_rel(Brain) = 363%) relative bioavailability compared with oral administration, underscoring its potential as an intranasal delivery system for efficient CNS targeting. Full article

Background/Objectives: PDE4 is a key regulator of cAMP signaling and a clinically validated anti-inflammatory target; however, the use of PDE4 inhibitors is often limited by adverse effects such as nausea, vomiting, and diarrhea. The natural compound braylin was previously identified as a novel PDE4 inhibitor scaffold, exhibiting an IC₅₀ of 0.96 µM. Using the PDE4–braylin co-crystal structure, we conducted structure-based design and optimization to enhance its potency. Methods: A series of novel braylin derivatives was synthesized and characterized. Their inhibitory activities against PDE4D were evaluated via enzymatic assays, and binding thermodynamics were analyzed by isothermal titration calorimetry (ITC). Molecular modeling was used to predict binding modes, and anti-inflammatory effects were assessed in LPS-stimulated macrophages. Results: Structure-guided optimization yielded lead compound L27, which showed significantly improved PDE4D inhibition (IC₅₀ = 67 nM) and high-affinity binding (K_d = 45 nM) as confirmed by ITC. L27 also exhibited remarkable selectivity against PDE isoforms. Molecular simulations highlighted key interactions with Gln369 and hydrophobic residues in the PDE4 active site. In cellular assays, L27 dose-dependently suppressed LPS-induced inflammation in macrophages at non-cytotoxic concentrations with efficacy comparable to roflumilast. Conclusions: We developed L27, a potent and selective PDE4 inhibitor derived from natural braylin. It demonstrated promising in vitro anti-inflammatory activity and represents a valuable lead for further therapeutic development. Full article

Chronic kidney disease (CKD) is a progressive global health challenge. While empagliflozin, a selective SGLT2 inhibitor, is known to attenuate CKD progression through mechanisms beyond glycemic control, the precise molecular pathways remain incompletely characterized and warrant further investigation. This study employed an integrated network pharmacology and molecular docking approach to elucidate the multi-target mechanisms of empagliflozin in CKD. Initial evaluation demonstrated that empagliflozin exhibits favorable physicochemical properties, drug-likeness, and ADMET profiles, supporting its potential as an effective orally administered therapeutic option for CKD management. Network analysis identified 221 shared molecular targets between empagliflozin and CKD-associated genes. Topological analysis of the protein–protein interaction (PPI) network revealed ten critical hub proteins—GAPDH, IL6, EGFR, HSP90AA1, NFKB1, HSP90AB1, MTOR, MAPK3, IL2, and PIK3CA—which serve as key regulators in CKD pathophysiology. Gene Ontology and KEGG pathway enrichment analyses indicated that these shared targets are significantly involved in phosphorylation, signal transduction, and central signaling cascades associated with CKD progression, including the PI3K-Akt, FoxO, HIF-1, and AGE-RAGE pathways. Molecular docking simulations corroborated empagliflozin’s multi-target affinity, demonstrating particularly strong binding energies toward HSP90AB1 (−10.85 kcal/mol), MAPK3 (−9.46 kcal/mol), and EGFR (−9.38 kcal/mol). Empagliflozin maintained stable hydrogen bonding throughout the 200-ns molecular dynamics simulation, primarily with GLN18, GLU42, SER45, ASN46, ASN101, GLY130, and TYR134, underscoring its persistent and well-anchored interaction with HSP90AB1. Collectively, these findings provide crucial mechanistic insights, suggesting that empagliflozin might exerts therapeutic effects by modulating interconnected pathways regulating inflammation, oxidative stress, and metabolic homeostasis, thereby reinforcing its role as a comprehensive, multi-target therapeutic strategy for CKD management. Nonetheless, validation through in vitro experiments remains necessary. Full article

Overexpression of protein arginine methyltransferase 5 (PRMT5) is pivotal in MYC-driven primary medulloblastoma tumors, suggesting PRMT5 as a potential therapeutic target. JNJ, a potent PRMT5 inhibitor currently in clinical trials, notably for non-Hodgkin lymphoma and lung cancer, was evaluated in this study. We report a validated LC–MS/MS bioanalytical method for quantifying JNJ in plasma and tissue matrices. The method demonstrated acceptable sensitivity, selectivity, and robustness in accordance with regulatory guidelines. The assay was linear over the range 0.2–500 ng mL⁻¹ (r² = 0.99), with plasma recovery exceeding 84% using only 100 µL of sample. Precision (%RSD < 15%) and accuracy (~91–108%) were within acceptable limits. JNJ showed >94% plasma protein binding and moderate Caco-2 permeability (3.4 ± 0.4 × 10⁻⁶ cm s⁻¹). Hepatic intrinsic clearance was higher in mouse liver microsomes than in human (41 ± 19 vs. 7 ± 0.6 mL min⁻¹ kg⁻¹). Following oral dosing in mice (10 mg kg⁻¹), T_max was 30 min with a C_max of 2781 ± 1033 ng mL⁻¹. Oral bioavailability was low (15%). The validated method was successfully applied to in vitro and in vivo studies and will guide dosing in animal models of medulloblastoma. Full article

Background/Objectives: Since 2016, the mutation of isocitrate dehydrogenase 1 (mIDH1) enzymes has become a major molecular marker for glioma classification and diagnosis. Moreover, the recent success of the INDIGO clinical trial on AG-881 (vorasidenib^®), an aminotriazine-based mutated IDH1/2 inhibitor (IC₅₀ = 6 nM/12 nM), validated the need for noninvasive detection of mIDH1 in brain tumors. This work is based on developing a series of novel fluorinated analogues of AG-881 and evaluating their potential in mIDH1 PET detection. Methods: The analogues were tested for their potency and then the best candidate was radiofluorinated and used for in vitro cell uptake studies. Results: Six analogues (6–11) were designed and synthesized, but only compound 6 showed nanomolar inhibitory potency towards mIDH1 (IC₅₀ = 400 nM). Following successful radiofluorination, in vitro cell uptake studies showed no selective accumulation of [¹⁸F]6. Conclusions: This study highlights the critical impact of substituent positioning and halogen substitution within the pyridyl moiety on maintaining inhibitory potency. Further medicinal chemistry research is needed to develop an aminotriazine-based ¹⁸F-radiolabeled mIDH1 ligand. Full article

The increasing demand for sustainable plant protection products has intensified interest in microbial biocontrol agents (BCAs). This study aimed to evaluate the antifungal activity of selected Streptomyces, Bacillus, and Trichoderma asperellum strains against phytopathogenic fungi and to assess their potential as BCAs under in vitro conditions. The antifungal activity of ten Streptomyces strains was first evaluated against Botrytis cinerea, Colletotrichum salicis, Fusarium oxysporum, and F. graminearum using a dual-culture assay. All isolates exhibited antifungal activity, with Streptomyces venezuelae MSCL 350 showing the strongest inhibition. In addition, the antifungal activity of T. asperellum MSCL 309 and three Bacillus strains was assessed against twelve Fusarium spp. isolates obtained from oats. T. asperellum demonstrated broad-spectrum inhibition, with growth inhibition ranging from 44.6% to 78.4%, primarily due to soluble metabolites, while volatile compounds showed no significant effect. Among the other tested Bacillus strains, only Bacillus subtilis MSCL 1441 exhibited antifungal activity, inhibiting all tested isolates. These results demonstrate strong strain-dependent antifungal activity and highlight T. asperellum MSCL 309, S. venezuelae MSCL 350, and B. subtilis MSCL 1441 as promising candidates for the development of environmentally friendly biocontrol agents. Full article

Exopolysaccharides (EPSs) are emerging as sustainable polymers for biomedical hydrogels. Here, we report hydrogels from sulfated EPSs produced by Porphyridium cruentum and ionically crosslinked with Ca²⁺, Ce³⁺, or Cu²⁺ to generate tunable networks with bioactive potential. Rheological analysis showed viscoelastic behavior was primarily governed by cation nature and accessible binding site density, with diminishing gains above 2.5 wt% EPS and limited benefit beyond 10 wt% crosslinker. Ce³⁺ produced the most solid-like gel, Ca²⁺ yielded more thixotropic networks, and Cu²⁺ promoted rapid, heterogeneous crosslinking consistent with fast surface complexation. These network signatures showed distinct in vitro performances. Cation selection tuned antibacterial activity against Staphylococcus aureus and Escherichia coli, with Cu²⁺ achieving rapid bactericidal effects and Ce³⁺ enabling an 8-log reduction after 24 h. The ABTS assay showed that Ca²⁺- and Ce³⁺-crosslinked gels had antioxidant potential (≥40 µM Trolox eq.mg⁻¹); however, antioxidant capacity was assay dependent. Conditioned-medium assays showed ≥75% viability at day 3 for Ca²⁺- and Ce³⁺-crosslinked gels against human dermal fibroblasts (HDFs), while only Ce³⁺-crosslinked gels were cytocompatible against human keratinocytes (HaCaTs). Cu²⁺-crosslinked gels were highly cytotoxic across all tested conditions. Macrophage cytokine readouts (TNF-α and IL-6) indicated formulation-dependent immunobiological response. This work establishes microalgal EPSs as versatile polymers and links crosslinking chemistry to rheological modulation and multifunctional biomedical performance, while direct wound-healing efficacy remains to be demonstrated in future in vivo or wound repair functional models. Full article

This study investigates the antioxidant properties of several synthetic peptides derived from milk proteins previously identified in milk permeate, a by-product of the dairy industry. The aim of the research is to identify which peptides present in milk permeate are responsible for its antioxidant activity. A comprehensive experimental strategy was employed to evaluate their antioxidant potential, including in silico selection, in vitro free radical scavenging assays and cellular models using Caco-2 and HCT116 cell lines. The peptides were screened using a molecular docking approach for their potential interaction with the Kelch-like ECH-associated protein 1/nuclear factor erythroid 2-related factor 2 (Keap1/Nrf2) pathway, and eight out of twenty-eight were selected and synthesized for further analyses. In vitro, six of the selected peptides demonstrated significant direct antioxidant activity in the DPPH scavenging assay, and two in the ABTS scavenging test. In cellular environments, three peptides (LPAPELGPRQA, LPIIQKLEPQI and NGQVWEESLKRL) effectively protect cells from oxidative stress induced by tert-butyl hydroperoxide, reducing reactive oxygen species production and partially mitigating lipid peroxidation. Further investigation showed that two of them (LPAPELGPRQA and LPIIQKLEPQI) effectively induce the Keap1/Nrf2 pathway, as evidenced by a ∼1.5-fold increase in Nrf2 levels and overexpression of downstream proteins. Permeability studies revealed that these peptides can cross the intestinal monolayer (2–3% in 2 h), suggesting potential systemic effects. Overall, these findings highlight the multifunctional antioxidant properties of the investigated peptides and support their potential application as nutraceuticals or therapeutic agents for oxidative stress-related conditions. Full article

In pancreatic cancer, cancer stem-like cells (CSCs) contribute to tumor initiation, reduced drug sensitivity, and recurrence. Limited strategies are currently available to target this cell population. Here we used a proteasome-low CSC enrichment system to identify microRNAs that negatively regulate CSC-like properties. From PANC-1 cells expressing a ZsGreen–ODC degron reporter, a proteasome-low population was isolated through sequential fluorescence-activated cell sorting of ZsGreen-positive cells. Molecular and functional analyses confirmed the CSC-like phenotype of this cell population. Integrated in silico analysis was used to select 31 microRNAs predicted to target CSC-related molecules, which were then evaluated by in vitro viability-based screening to identify candidates that selectively suppressed the viability of CSC-like cells, relative to non-CSCs. Moreover, comprehensive miRNA expression profiling revealed that miR-136-5p was downregulated in the CSC-like population and was therefore selected for further analysis. Mechanistically, miR-136-5p directly targets the 3′ untranslated region of DCLK1 and reduces its expression, with a greater reduction in the short isoform. Finally, in a CSC-derived xenograft mouse model, systemic delivery of miR-136-5p using super carbonate apatite nanoparticles significantly suppressed tumor growth. Taken together, these findings suggest that miR-136-5p restoration may provide a therapeutic approach for targeting CSC-driven tumor growth in pancreatic cancer. Full article

The hypoxic microenvironment plays a critical role in the progression of canine osteosarcoma (OSA) by promoting different cellular responses, including the release of extracellular vesicles (EVs). Given the clinical aggressiveness of canine OSA, the aim of this study was to evaluate the miRNAome profile in EVs released in vitro by four canine OSA cell lines under hypoxic conditions. In particular, for this study we used two commercial canine osteosarcoma cell lines (D17 and D22) and two primary osteosarcoma cell lines obtained in our laboratory (Penny and Wall). D17, D22, Penny, and Wall cell lines were cultured under normoxic and hypoxic conditions (200 µM CoCl₂) for 24 h. EVs were isolated by size-exclusion chromatography and characterized by nanoparticle tracking analysis and Western blotting. miRNAs extracted from EVs were then sequenced and analyzed using bioinformatics approaches. The most representative miRNAs were identified and validated by qPCR using the miRCURY LNA miRNA PCR assay. miRNome profiling identified 233 miRNAs differentially expressed in EVs across all analyzed cell lines. Among these, 94 miRNAs were detected exclusively under hypoxic conditions. From this subset, 43 miRNAs were selected for further validation by qPCR. The qPCR results showed that miR-222-3p and miR-186-5p were significantly downregulated in the Wall cell line under hypoxia (p ≤ 0.05). TargetScan and pathway enrichment analyses demonstrated that miR-186-5p regulates target genes involved in different cellular processes. In human osteosarcoma, low serum levels of miR-222-3p are associated with poor prognosis, while miR-186-5p is recognized as a key hypoxia-responsive miRNA. Collectively, these results suggest the potential of EV-associated miRNAs as biomarkers in canine OSA and support their relevance in translational and comparative oncology. Full article

Background: The wear resistance of modern commercial glass-ceramic materials used in dental prostheses was investigated under cyclic contact conditions that included a rotational component. This loading mode has been largely overlooked in conventional in vitro wear testing, yet may be clinically relevant in patients with parafunctional conditions such as bruxism. Methods: Rotational loading was applied using an all-electric testing machine equipped with a biaxial actuator. Loading cycles combined a normal load (50 N) and a rotation (30°), at a frequency of 1 Hz. Microstructure and damage were characterized using advanced microscopy. Results: Rotational loading induced substantial damage across this class of materials, including the formation of glassy tribolayers with limited protective capability under the low-humidity conditions examined. Significant microstructure-dependent variations in wear volume were observed, with specific wear rates indicating severe wear (SWR above 10⁻⁶ mm³/N·m threshold) in three of the five materials tested. Lithium disilicate glass-ceramics, characterized by a high fraction of elongated reinforcement crystals, exhibited the greatest resistance to damage, whereas leucite-based glass-ceramics showed the lowest. The dominant wear mechanisms were plastic-deformation-induced grooving and fracture-driven chipping. The findings are interpreted within established wear models for brittle materials (Archard and fracture-based) and supported by numerical simulations of stress fields across multiple length scales. Implications: The results provide mechanistic insight into rotational wear damage in glass-ceramic systems, a material class particularly susceptible to such loading, and inform strategies for material selection and microstructural design aimed at improving prosthetic durability. Full article

Background: Electrochemotherapy enhances the intracellular delivery of anticancer agents through electroporation but is traditionally limited to cytotoxic drugs associated with significant side effects. Vitamin C (ascorbic acid) exhibits selective anticancer activity when accumulated at high intracellular concentrations; however, its therapeutic application is restricted by poor membrane permeability and rapid systemic clearance. Methods: In this study, we investigated whether reversible electroporation, applied using a custom-designed variable plate electrode system designed to deliver a uniform electric field, could potentiate the antitumor efficacy of low-dose vitamin C. Numerical simulations were performed to optimize electrode spacing and stimulation voltage, suggesting homogeneous electric field coverage throughout the tumor volume. The proposed approach was evaluated in vitro using MDA-MB-231 and 4T1 breast cancer cell lines and in vivo in a 4T1 murine breast cancer model. Results: Low-dose vitamin C alone produced minimal cytotoxic effects, whereas its combination with electroporation significantly reduced cell viability and increased apoptotic and necrotic cell death in vitro. In vivo, vitamin C–assisted electrochemotherapy resulted in pronounced tumor growth suppression, with tumor volumes reduced to approximately 0.34-fold of baseline by day 15, accompanied by decreased proliferation and marked tissue disruption. Conclusions: These findings demonstrate that uniform-field reversible electroporation markedly enhances the intracellular delivery and antitumor activity of low-dose vitamin C, supporting this technology-driven strategy as a promising, low-toxicity alternative to conventional chemotherapeutic agents in electrochemotherapy for solid tumors. Full article

Lycium barbarum–Polygonatum cyrtonema composite jiaosu (LBPCJ) was prepared by sequential dual-species fermentation and evaluated in a mouse model of alcohol-induced liver injury. Following process optimization, a yeast-first sequential strategy with intermediate pasteurization was selected, comprising an initial Saccharomyces cerevisiae fermentation step, intermediate pasteurization, and a subsequent Lactiplantibacillus plantarum fermentation step. Fermentation reduced pH from 4.68 to 3.51 and increased total acidity from 61.06 to 135.39 g LA/L and total phenolic content from 3.01 to 9.39 mg GAE/mL. In vitro antioxidant-related activities were also higher after fermentation, with DPPH, ABTS, and •OH scavenging rates increasing by 39.90%, 29.78%, and 11.10%, respectively. In mice, LBPCJ administration was associated with lower liver index and serum aminotransferase levels, together with attenuated hepatic histopathological alterations, with the high-dose group (15 mL/kg BW) showing the clearest response. These changes were accompanied by higher hepatic SOD and GSH levels and lower MDA, TNF-α, IL-1β, and IL-6 levels. LBJ and PCJ also improved several measured indicators, while LBPCJ showed changes across multiple endpoints under the tested conditions. Overall, sequential fermentation markedly altered the physicochemical and antioxidant-related properties of LBPCJ, and LBPCJ administration improved multiple indicators related to alcohol-induced liver injury in mice, although the specific constituents and underlying mechanisms remain to be clarified. Full article

Macrophages are highly plastic cells of the tumor microenvironment, and although classically activated M1 macrophages are generally regarded as anti-tumor effectors, their prolonged cytotoxic activity may also promote tumor adaptation. In this study, we investigated whether sustained exposure of prostate cancer cells to cytotoxic M1-like macrophages results in the selection of tumor cell populations with enhanced malignant properties. Macrophage-resistant derivatives of the human prostate cancer cell lines PC3 and DU145 were generated by repeated co-culture with cytotoxic THP-1-derived macrophages and characterized in vitro and in vivo. Macrophage-selected tumor cells showed increased proliferative activity and enhanced clonogenic survival. In vivo, these cells formed larger xenograft tumors with more aggressive histopathological features. At the same time, their migratory activity was not significantly increased, although they displayed partial epithelial–mesenchymal transition-like changes, including increased vimentin expression without a marked loss of epithelial markers. Transcriptomic profiling revealed coordinated inflammatory and adaptive reprogramming, characterized by the enrichment of cytokine- and immune-response pathways together with the suppression of metabolic and differentiation-associated programs. These changes were accompanied by the selective activation of p38 MAPK signaling, while sensitivity to paclitaxel remained unchanged. Taken together, the results indicate that macrophage cytotoxicity may act as a selective pressure promoting the emergence of inflammation-adapted tumor cell variants with increased malignant potential, supporting re-evaluation of the role of M1-like macrophages in tumor progression. Full article