Acta Pharmacologica Sinica最新文献

Serotonin-gated 5-HT3 receptors (5-HT3Rs), which belong to the cys-loop superfamily of ligand-gated ion channels, mediate fast excitatory neurotransmission in the central and peripheral nervous system. They are targets for drugs to treat neurological diseases and psychiatric disorders, as well as chemotherapy-induced and postoperative nausea and emesis. The ever-increasing number of resolved 3D structures of the homopentameric form of 5-HT_3AR, in combination with new computational approaches allow us to better understand the molecular processes of ligand binding, the subsequent conformational changes, and ion permeation, providing a solid foundation for understanding the biological functions of 5-HT_3AR and its heteropentameric 5-HT3R homologs. In this review, we first outline the physiological roles and subunit assembly of heteromeric 5-HT3Rs, which predominate in vivo. We then summarize the latest structural insights into the 5-HT_3AR, revealing details of its architecture, ligand-binding sites, and conformational transitions leading to channel activation. Finally, we discuss the evolving pharmacology of 5-HT3R modulators and provide our perspectives on future research directions aimed at resolving the heteropentameric structures of 5-HT3R in their native membrane and developing modern drugs targeting these receptors.

Autoantibody- and complement-mediated cytotoxicity can cause autoimmune astrocytopathy that leads to CNS inflammatory demyelination. Formyl peptide receptor 2 (FPR2/ALX) governs the activation and propagation of immune response. However, the precise role of FPR2/ALX in neuroinflammation and the effect of FPR2/ALX stimulation on autoimmune astrocytopathy are poorly understood. Using a mouse model of autoimmune astrocytopathy induced by AQP4-IgG- and complement-mediated cytotoxicity, we found that the stimulation of FPR2/ALX with the small-molecule agonist Quin-C1 led to reduced brain lesion volume, astrocyte loss and demyelination. This was accompanied by enhanced anti-inflammatory activity of microglia and reduced infiltration of lymphocytes in the brain. FPR2/ALX stimulation also led to increased phosphorylation of SYK and AKT in mice with autoimmune astrocytopathy. Notably, the benefits of FPR2/ALX stimulation were attenuated in mice with autoimmune astrocytopathy after microglial depletion using the CSF1R inhibitor PLX5622 or natural killer (NK) cell depletion using an anti-NK1.1 monoclonal antibody. Additionally, the protective effects of FPR2/ALX stimulation were diminished in mice with autoimmune astrocytopathy that received the SYK inhibitor R406. Collectively, our findings demonstrate that FPR2/ALX stimulation may represent a promising therapeutic strategy to attenuate detrimental neuroinflammation in autoimmune astrocytopathy by modulating microglia and NK cells. FPR2/ALX stimulation suppresses autoimmune astrocytopathy: Using a mouse model of autoimmune astrocytopathy, we demonstrated that FPR2/ALX stimulation with the small molecule Quin-C1 reduces the CNS infiltration of lymphocytes and augments the anti-inflammatory activity of microglia, leading to attenuated astrocyte pathology induced by AQP4-IgG and complement-mediated attacks. Mechanistically, the benefits of FPR2/ALX stimulation using Quin-C1 involve microglia, natural killer (NK) cells, and SYK-AKT signaling.

Peroxiredoxin 1 (PRDX1) is a pivotal antioxidant enzyme maintaining intracellular reactive oxygen species (ROS) balance. Deficiency of PRDX1 aggravates oxidative stress-related pathologies, whereas enhanced PRDX1 activity confers cytoprotection. Small-molecule agonists boosting PRDX1 peroxidase activity hold therapeutic promise, yet to date only two such agonists-rosmarinic acid (RA) and salvianolic acid B (SAB)-have been reported, both by our laboratory. These polyphenolic compounds are chemically rigid and recalcitrant to modification. Here, we resolved the crystal structure of PRDX1 in complex with salvianolic acid C (SAC), revealing a conserved danshensu substructure shared by SAC, RA, and SAB. Guided by this pharmacophore, we designed a scaffold hopping core structure and generated 160 derivatives via in situ click reaction. Among them, LC-PDA-01, a non-polyphenolic scaffold, exhibited the highest PRDX1 activation (EC₅₀ = 111.8 nM). This work discloses the first structurally tractable PRDX1 agonist and highlights combinatorial click chemistry's utility in transforming natural product motifs into drug-like molecules.

Radiopharmaceutical therapy (RPT) represents a critical approach in oncology, nevertheless its efficacy may be limited by tumor resistance mechanisms associated with metabolic reprogramming. Enhancing tumor radiosensitivity remains a major challenge. Engineered bi-functional starvation probes (CRT3LP and CRT4LP) that simultaneously target ecto-CRT and exert L-ASNase activity are explored for disrupting tumor amino acid metabolism. Herein, we systematically evaluate the ability of targeted starvation probes to enhance antitumor efficacy in radioactive iodine (RAI) therapy. In vitro, the probes upregulated p53 expression while downregulating Rev1 and SOD₂, thereby impairing ROS scavenging and sensitizing tumor cells to RAI-induced oxidative stress. In vivo, the combination treatment elevated intratumoral ROS levels, increased CD4⁺ and CD8⁺ T cell infiltration, upregulated pro-inflammatory cytokines (IFN-γ and TNF-α), and reduced regulatory T cell populations. Additionally, markers of tumor proliferation (Ki67 and CD31) were suppressed, while apoptotic markers (TUNEL and p21) were increased. Co-administration of the immune checkpoint inhibitor αPD-L1 further improved therapeutic efficacy. These findings suggest that targeted tumor starvation probes boost radiosensitivity and anti-tumor immunity, and this strategy shows improved efficacy in combination with αPD-L1 therapy.

Recent advances in structural biology, functional genomics, and artificial intelligence (AI) have expanded understanding of the solute carrier (SLC) transporter superfamily. Emerging evidence indicates that SLC transporters play central roles in the multi-hit pathogenesis of metabolic dysfunction-associated fatty liver disease (MAFLD). In this review, we build upon the classical "two-hit" hypothesis to summarize how CRISPR-based screening, cryo-electron microscopy (cryo-EM), and AI-driven platforms such as AlphaFold and RosettaVS have advanced the study of SLC transporters. These approaches have enabled the identification of SLCs implicated in MAFLD pathogenesis, including SLC13A5 (NaCT) and SLC25A47, as well as the determination of high-resolution structures that support rational drug design for targets such as sodium-glucose cotransporter 2 (SLC5A2) and monocarboxylate transporter 10 (SLC16A10). In addition, ultra-large virtual screening strategies have accelerated the discovery of small-molecule inhibitors targeting SLC transporters. We synthesize current evidence defining the mechanistic roles of SLC transporters in lipid and glucose metabolism, mitochondrial dysfunction, and gut-liver axis dysregulation. Finally, we discuss therapeutic implications, ranging from clinically repurposed SGLT2 inhibitors to investigational agents such as the SLC13A5 inhibitor ETG5773, and outline future directions for technology-driven precision medicine treating MAFLD.

Biomolecular condensates formed through liquid-liquid phase separation (LLPS) have emerged as central organizers of cellular biochemistry. In cancer, dysregulated phase separation gives rise to oncogenic condensates that reprogram transcriptional, signaling, and stress-response networks. By selectively concentrating oncogenic proteins and nucleic acids, these dynamic assemblies create permissive regulatory microenvironments that stabilize malignant cell states and facilitate therapeutic resistance. Accumulating evidence now indicates that oncogenic condensates are pharmacologically tractable, opening new opportunities for targeted intervention. In this Review, we synthesize current mechanistic insights into condensate assembly, regulation, and material state transitions; examine their context-dependent behaviors and functional heterogeneity across tumor ecosystems; and delineate emerging therapeutic strategies, including small-molecule modulators, peptide-based inhibitors, targeted protein degraders, and RNA-directed approaches. We further highlight recent translational advances-exemplified by DPTX-3186, a first-in-class condensate modulator granted FDA Orphan Drug Designation for Wnt-driven gastric cancer-that underscore the clinical promise of condensate-directed therapies. Finally, we discuss key mechanistic, pharmacodynamic, and biomarker-related challenges that must be addressed to fully integrate condensate biology into next-generation precision oncology.

Despite the increasing recognition of pyroptosis, particularly that involving GSDME, its precise impact on tumor prognosis and the immune microenvironment remains elusive, necessitating a comprehensive investigation in the context of lung adenocarcinoma (LUAD). We aimed to construct a pyroptosis-related prognostic model and to elucidate the intricate dynamics of GSDME-mediated pyroptosis in shaping tumor immunity in LUAD. We developed a pyroptosis-related prognostic model using machine learning. GSDME-mediated pyroptosis in LUAD cells was induced using CHX and TNF-α. HMGB1 content in the cell supernatant after cell pyroptosis and in serum from patients before treatment with PD-1/PD-L1 antibodies was determined by Enzyme-Linked Immunosorbent Assay. In vivo, Lewis lung carcinoma (LLC)-bearing C57 mice were treated with cisplatin and/or caspase-3 inhibitors, anti-PD-1, and IL-8 inhibitors, with tumor growth monitored. Our prognostic prediction model (PYR_score), built upon pyroptosis-related genes, demonstrated high efficacy in predicting LUAD prognosis across diverse datasets. Machine learning analyses revealed that higher PYR_score values correlated with shorter progression-free and overall survival. CHX and TNF-α induced GSDME-mediated pyroptosis with elevated HMGB1. Increased HMGB1 was associated with worse therapeutic efficacy of immune checkpoint inhibitors in LUAD patients. HMGB1 increased the proliferative ability and IL-8 secretion of Treg cells in vitro. Caspase-3 and IL-8 inhibitors slowed tumor growth, and IL-8 inhibitors possibly enhanced the effectiveness of anti-PD-1 immunotherapy in LLC-bearing mice. In summary, our novel PYR_score is a robust prognostic marker, offering predictive power across different datasets. GSDME-mediated pyroptosis modulated the immunosuppressive microenvironment via elevations in HMGB1, Treg cells, and MDSCs. IL-8 inhibitors may inhibit Tregs and MDSCs and enhance the effectiveness of anti-PD-1 immunotherapy. Further clinical validation and exploration of therapeutic interventions targeting these pathways are essential for translating these findings into clinical practice.

Several essential physiological systems express voltage-gated potassium channels within the K_V7 family (comprising K_V7.1-7.5), sometimes also co-assembled with auxiliary subunits in the KCNE family (comprising KCNE1-5). An ongoing challenge to K_V7 drug development is creating subtype-selective compounds to limit adverse effects. Prior work has shown that the antiepileptic cannabidiol (CBD), a pan-K_V7 modulator, inhibits cardiac- and epithelia-associated K_V7.1 and K_V7.1/KCNE1 channels, while activating neuronal K_V7 subtypes (K_V7.2-7.5). However, little is known about the binding sites through which CBD mediates inhibitory effects on K_V7.1 and K_V7.1/KCNE1, limiting insight towards the development of selective K_V7 modulators. To address this knowledge gap, we used a combination of the Chai-1 artificial intelligence model (to generate CBD binding site predictions in human K_V7.1 and K_V7.1/KCNE1 channels), site-directed mutagenesis and electrophysiology of these channels expressed in Xenopus laevis oocytes (to corroborate CBD binding site predictions), and molecular dynamics simulations (to study the biophysical mechanisms underlying CBD binding). We found that CBD binds to two unique sites within K_V7.1 and K_V7.1/KCNE1. In K_V7.1 alone, CBD was bound to an intrasubunit S5-S6 pore domain binding site; referred to as the S5-S6 site. In K_V7.1/KCNE1, the addition of the KCNE1 subunit created a novel binding site for CBD, sandwiched between two K_V7.1 subunits and one KCNE1 subunit; referred to here as the S6-S5'-E1 site. Molecular dynamics simulations showed that CBD binding to the S6-S5'-E1 K_V7.1/KCNE1 site closes off the K_V7.1 S5-S6 site. A sequence comparison between K_V7 channels revealed key amino acid differences at both the S5-S6 and S6-S5'-E1 sites relative to neuronal K_V7s. These support the notion that CBD binds differently in K_V7.1 and K_V7.1/KCNE1 channels in accordance with its unique inhibitory pharmacological effects on these channels compared to the activating effect in neuronal K_V7s. Thus, we provide support for K_V7.1 and K_V7.1/KCNE1 being inhibited by CBD via distinct binding sites, which can guide future research focused on the rational development of drugs that avoid inhibitory effects on K_V7.1 and K_V7.1/KCNE1 channels or utilize these sites to modulate channel activity.