Journal of pharmaceutical analysis最新文献

Catechol-O-methyltransferase (COMT) inhibition is a critical therapeutic strategy for Parkinson's disease (PD), yet clinical inhibitors face limitations in bioavailability and hepatotoxicity, driving demand for novel natural scaffolds. In this study, we developed an integrated analytical platform by coupling high-resolution bioassay profiling (HRBP) and affinity-based ligand fishing system to effectively characterize bioactive compounds targeting COMT in Paeonia lactiflora Pall. (the most frequently used core herb in tradition Chinese medicine prescriptions for PD treatment). Parallel high-performance liquid chromatography with diode-array detection and tandem mass spectrometry (HPLC-DAD-MS/MS) coupled with nanofractionation enabled real-time bioactivity mapping via 384-well COMT inhibition assays, while semi-preparative liquid chromatography (LC) was employed to further identify co-eluted components. HRBP and immobilized COMT ligand fishing identified 16 and 21 candidates, respectively, with 5 overlapping bioactive markers. Notably, the potent inhibitors galloylpaeoniflorin (half maximal inhibitory concentration (IC₅₀) = 16.2 ± 3.4 μM) and 1,2,3,4,6-O-pentagalloylglucose (IC₅₀ = 3.1 ± 0.5 μM) exhibited comparable potency to the positive control morin (IC₅₀ = 10.1 ± 0.7 μM). Molecular docking results further revealed the critical interactions and binding sites between the active compounds and COMT. The validated platform demonstrates significant potential for rapid discovery of plant-derived enzyme inhibitors, bridging advanced separation, bioactivity screening, and mechanistic validation in neurodegenerative therapeutic development.

This study explores the molecular mechanisms behind the remote transfer of thyroid cancer (THCA) by investigating the interaction network of C-X-C motif chemokine ligand 8⁺ (CXCL8⁺ monocytes and syndecan-1⁺ (SDC1⁺) tumor stem cells using single-cell and spatial transcriptome sequencing. Tumor samples from THCA patients were analyzed using single-cell RNA sequencing (scRNA-seq), spatial transcriptome sequencing, and tumor tissue transcriptome analysis. Data were processed with Seurat and CellChat R packages, integrated via the SPOTlight package, and correlated with clinical data from the UCSC Xena database. Functional pathway enrichment analyses were performed using Gene Set Enrichment Analysis (GSEA), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genome (KEGG). In vitro, a co-culture system of monocytes and THCA stem cells was developed, and protein levels were measured via enzyme-linked immunosorbent assay (ELISA) and Western blotting. The self-renewal and migration of follicular thyroid carcinoma (FTC) 238-S cells were assessed through sphere formation, colony formation, Cell Counting Kit-8 (CCK-8), and Transwell assays. In vivo, a subcutaneous tumor xenograft model and a lung metastasis model were established in nude mice. Transcriptomic analyses identified the CXCL8/SDC1 axis as a key mediator of Janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling activation, promoting THCA stem cell self-renewal, invasion, and metastasis. CXCL8/SDC1 expression was significantly higher in the high-risk C1 subtype of THCA patients and correlated with a worse prognosis. In vitro and animal studies confirmed that the CXCL8/SDC1 axis drives tumor progression and metastasis. The interaction between CXCL8⁺ monocytes and SDC1⁺ tumor stem cells activates the JAK-STAT pathway, facilitating the remote transfer of THCA. Targeting the CXCL8/SDC1 axis may provide novel therapeutic strategies for improving THCA patient outcomes.

Depression is a prevalent mental disorder characterized by persistent disinterest and a depressed mood, with severe cases potentially leading to suicide. In recent years, the incidence of depression has steadily increased, making it the second-largest global health burden. The pathogenesis of depression involves a series of complex pathological mechanisms, although the key underlying causes remain unclear. Programmed cell death (PCD), including apoptosis, autophagy, pyroptosis, ferroptosis, and necroptosis, involves highly organized gene expression processes that may influence the occurrence and development of depression by regulating cellular fate. Furthermore, numerous studies have shown that natural products can modulate PCDs through various signaling pathways, presenting significant potential for managing depression. Natural products offer benefits such as cost-effectiveness, fewer side effects, and other advantages, making them viable supplements or alternatives to traditional antidepressant drugs. To explore this potential, we reviewed studies demonstrating the antidepressant effects of natural products through multi-target modulation of PCDs. In addition, we discussed the toxicity and clinical applications of these natural products. This study highlights that diverse core biological pathways and targets are involved in determining the fate of depression-associated brain cells, including the PI3K/Akt signaling pathway, caspase-8, GSDMD, and others. In conclusion, the multi-target mechanisms of PCD regulation by natural products may provide a promising foundation for the future development of novel antidepressant medications.

The emergence of drug-resistant bacterial infection and persistent biofilm colonization pose a rigorous challenge to effective wound healing and regeneration, necessitating the innovative therapeutic strategies to combat these pressing clinical crises. Herein, nortriptyline, a novel United States Food and Drug Administration (FDA)-approved tricyclic antidepressant was uncovered to effectively potentiate bactericidal activities of β-lactam antibiotics against methicillin-resistant Staphylococcus aureus (MRSA). Mechanistically, nortriptyline functions by disrupting the microbial iron homeostasis and potentiation of Fenton chemistry-mediated oxidative stress, concomitant with metabolic reprogramming via tricarboxylic acid (TCA) cycle dysregulation and membrane destabilization. To enhance combination therapy-mediated therapeutic potential in wound management, the dual-loaded self-healing hydrogel OHA-PLL@AN was engineered to exhibit excellent biocompatibility and antibacterial potentials through molecular cross-linking of oxidized hyaluronic acid (OHA) and ε-polylysine (PPL). The therapeutic efficacy of OHA-PLL@AN was further validated in a murine model with MRSA-infected cutaneous wounds. OHA-PLL@AN therapy significantly attenuated the inflammatory response, concurrently promoting angiogenesis and accelerating the cutaneous wounds healing. Collectively, these findings underscore the dual drug-loaded self-healing hydrogel OHA-PLL@AN with anti-infection and anti-inflammatory properties as a novel therapeutic strategy for drug-resistant bacterial infected wounds therapy.

[This corrects the article DOI: 10.1016/j.jpha.2022.10.002.].

Anthocyanins, a unique class of flavonoids with flavylium skeletons, are valued for antioxidant properties. However, distinguishing anthocyanins from co-existing flavonoids using conventional automated tandem mass spectrometry (MS) analysis methods remains challenging. This difficulty arises from low specificity of MS features and confusion of precursor ions, leading to substantial false confidence annotations. To address it, we have developed the strategy of positive (POS)-to-negative (NEG) primary MS (MS¹) intensity ratios detecting with fast polarity switching (FPS), termed FPS-POS/NEG, to determine their specific precursor ions. Moreover, we developed an automated program leveraging FPS-POS/NEG strategy (FPS_P/N) streamlining screening candidate pool with molecular networking analysis from MS¹ and secondary MS (MS²), determining precursor ions with FPS-POS/NEG, and annotation with MS². This program enables simultaneous capture of positive and negative signals in a single run and accurate determination of precursor ions for anthocyanins (5.98-9.28) and other flavonoids (-2.52 to 2.08). The underlying mechanisms were elucidated by difference in protonated and deprotonated Gibbs free energy (ΔG) and in-source fragmentation (ISF). FPS-POS/NEG strategy was validated across a broad pH range (0.1%-2% formic acid (FA)) and demonstrated high alignment accuracy (retention time difference, 0.011 min) and consistency (relative standard deviation (RSD), 0.38%-4.62%). Using blueberry, 20 anthocyanins (nonacylated and acylated) and 14 additional flavonoids were annotated. With two-dimensional integration of positive and negative MS¹ intensities with intensity ratios, FPS_P/N program provides a novel way to identify anthocyanins from other flavonoids. We anticipate this innovative method will enhance the high-throughput qualification of anthocyanins and other flavonoids in complex samples.

The cuproptosis-related gene (CRG) SH3GL1 is identified as a pivotal regulator in breast cancer (BRCA) progression and immune regulation in this study. Through gene expression profiling and meta-analysis of public datasets, SH3GL1 was found to be overexpressed in BRCA tumor tissues and correlated with poor prognosis. Single-cell RNA sequencing pinpointed SH3GL1's expression in epithelial cells and its critical interactions with immune cells, particularly T cells and monocytes. Functional experiments confirmed SH3GL1's role in promoting immune cell migration and modulating drug sensitivity. Moreover, high SH3GL1 expression was linked to reduced immunotherapy response, as revealed by TIDE scoring, suggesting its contribution to the immune microenvironment complexity in high-risk BRCA groups. These results emphasize SH3GL1's dual role as a prognostic biomarker and a target for therapeutic intervention in BRCA, providing new insights into personalized cancer treatment approaches.

While being a safe and effective precision therapy strategy, tumor immunotherapy still fails in many patients due to the immunosuppressive tumor microenvironment. Emerging evidence has indicated that the targeted nano-drug delivery systems can accurately deliver therapeutic agents to potentiate the efficacy of immunotherapy. This review will outline recent advances in applying targeted nano-drug delivery systems in immunotherapy, with an emphasis on their crucial roles in regulating innate immune responses, adaptive immune responses, and immunogenic cell death. We will also discuss the current challenges and future opportunities for the clinical translation of targeted nano-drug delivery systems for tumor immunotherapy.

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Hepatocellular carcinoma (HCC) is the predominant type of liver cancer. There are different risk factors for HCC including viral infection, liver fibrosis, non-alcoholic fatty liver disease, environmental factors and genomic alterations. The tumor microenvironment (TME) has been proposed as a potent regulator of tumor malignancy comprised of normal and cancerous cells. Macrophages are among the most abundant cells in the TME, known as tumor-associated macrophages (TAMs) that can control proliferation, metastasis, immune reactions and therapy response of tumor cells. In the present review, the function of TAMs in the regulation of HCC progression was evaluated. TAMs are prognostic factors in HCC that increase in TAM infiltration into TME can cause undesirable outcome in patients. Moreover, M2 polarization of macrophages can impair function of other immune cells such as T cells and natural killer (NK) cells to mediate immune evasion. TAMs demonstrate association with other biological events including autophagy and glycolysis. There is mutual interaction between TAMs and exosomes that TAM-mediated exosome secretion regulates HCC progression, while exosomes derived from other cells can also affect TAMs. Inhibition of macrophage recruitment, their depletion and increasing M1 polarization are promising approaches in HCC therapy. The natural products and nanostructures have been also recently introduced for the regulation of macrophages in HCC therapy.