ACS Pharmacology and Translational Science最新文献

Xanthone, an oxygenated heterotricyclic phytochemical, has recently emerged as a promising scaffold in cancer research due to its multitargeted anticancer potential. Recent studies demonstrate its affinity for G-quadruplex (G4) DNA structures. In this context, herein, we describe the design and synthesis of a series of xanthone–benzimidazole conjugates to investigate their specific cytotoxic effects on cancer cells through the stabilization of telomeric G4 DNA structures. Comprehensive in vitro biophysical and cellular experiments were performed to screen the most effective compounds for targeting telomeric G4 DNA structures among them. The structure–activity relationship (SAR) of quadruplex-compound interactions was documented as well. The current study reveals that two compounds, XDBHEP and XDBAEP, exhibit maximum binding affinity and selectivity toward the antiparallel telomeric form of G4 DNA. These compounds stabilize this G4 DNA structure through binding within the DNA groove loci and exhibit a 1:1 molar binding stoichiometry. The specific cytotoxic effect of these compounds on different types of cancer cells, including A549, T-47D, and MCF-7, and their apoptotic-mediated cell death was further demonstrated by several in vitro cellular experiments. In addition, blood compatibility, ADME studies, pharmacokinetics, and biodistribution studies were also performed to assess the potential of these compounds for further in vivo and clinical investigations. Based on the current study, the selective antiparallel telomeric G4 DNA targeting properties and specific cancer cell cytotoxicity effects of xanthone–benzimidazole conjugates could provide valuable information to support the research community in the future development of new anticancer drugs through G4 DNA stabilization based on this pharmacophore.

Viral proteases are critical targets for antiviral drug development, but current screening methods for protease inhibitors often require high biosafety levels or lack cell-based relevance. Here, we developed a novel cell-based assay system utilizing recombinant green fluorescent protein (GFP) technology, designated as DIFF-recombinant GFP (DIFF-rGFP), for potentially high-throughput screening of viral protease inhibitors. By systematically investigating potential insertion sites within the green fluorescent protein (GFP), we constructed a series of recombinant green fluorescent proteins (rGFPs) that accommodate exogenous protease cleavage sequences. Using the 3-Chymotrypsin like protease (3CL^pro) of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) as a model, we demonstrated that the DIFF-rGFP assay relies on the coexpression of rGFP and the protease, with fluorescence intensity increasing upon inhibitor action. This assay eliminates the need for high biosafety laboratories and is performed at the cellular level. For proof of concept, we validated this method using two well-characterized SARS-CoV-2 3CL^pro inhibitors, GC376 and ensitrelvir, to demonstrate its applicability for inhibitor screening. Our results indicate that the DIFF-rGFP assay is a safe, efficient, and reliable platform for identifying viral protease inhibitors with potential applications in accelerating antiviral drug discovery.

This Viewpoint examines the emerging role of cannabidiol (CBD) in sports medicine, with a particular emphasis on its potential to support athlete health through indirect mechanisms. Rather than acting as a direct performance enhancer, CBD may contribute to improved readiness for training and competition by promoting better sleep, alleviating anxiety, and accelerating recovery. We also discuss challenges for antidoping compliance, including product contamination and regulatory gaps in Brazil, and highlight CBD’s promise as a safer alternative to opioids for pain management in athletes.

The circadian rhythms and cell cycle are closely interlinked, creating a fundamental regulatory axis vital for tissue homeostasis, which is frequently dysregulated in cancers. The circadian apparatus, which is regulated by the core clock components (BMAL1, CLOCK, PER, and CRY in mammals), establishes temporal order on cell proliferation by rhythmically regulating important cell cycle regulators such as WEE1, p21, and the oncogene MYC. This is frequently accomplished through overlapped signaling nodes that include particular kinases and ubiquitin ligases (e.g., FBXW7). Mounting evidence implicates disruption of this circadian clock-cell cycle synchrony, arising from genetic or environmental factors, as a significant contributor to tumorigenesis and progression via impacts on DNA repair fidelity, oncogene stability, and tumor suppressor pathways. This review critically evaluates the new concept of chrono-pharmacology for cancer, focusing on the substantial effects and side effects of different anticancer drugs that depend on the time-of-day efficacy. We discussed some interesting examples, like HSP90 inhibitors (ganetespib), HDAC inhibitors (quisinostat), topoisomerase inhibitors (doxorubicin), and BCL-2 family antagonists (Obatoclax, TW-37), whose therapeutic activities are tightly regulated by circadian control over their molecular targets, pharmacokinetic processes, and downstream physiological pathways. Furthermore, the circadian influence extends to the tumor microenvironment and antitumor immunity, suggesting novel chrono-immunotherapy approaches. By putting together the molecular bases of these temporal dynamics, this review underscores the significant potential of chronotherapy─the timed administration of drugs to improve cancer treatment by enhancing therapeutic indices and paving the way for personalized, temporally optimized oncology strategies.

Class A G protein-coupled receptors (GPCRs) are targets for ∼36% of commercial drugs. GPCRs in their apo-forms exhibit conformational heterogeneity, and more than a single active and inactive conformation exists in equilibrium. Distinct transient conformational states can be significantly populated and can be coupled with different agonists, transducers, and effectors, giving rise to divergent signaling pathways. The characterization of such transient conformational states, which may have eluded identification by X-ray crystallography and cryogenic electron microscopy, can be achieved through a combination of biophysical techniques, such as nuclear magnetic resonance, double electron–electron resonance spectroscopy, single-molecule fluorescence microscopy, molecular dynamics simulations, and mass spectrometry. We review findings about the functional, conformational states of four class A GPCRs, including detailed results for the adenosine A_2A and β₂ adrenergic receptors and important observations for the β₁ and μ opioid receptors. The identification of ligands that can bind to distinct conformations, e.g., agonists that activate favorable pathways while inhibiting deleterious ones, represents an important goal in drug development.

Leukemia Inhibitory Factor (LIF) is a pleiotropic cytokine secreted by tumor cells to evade immune detection, contributing to tumor progression and resistance to therapy. Targeting LIF has emerged as a promising strategy, with anti-LIF therapies in clinical trials across a variety of cancers. Glioblastoma, a highly aggressive LIF-secreting brain tumor, is a critical target for these emerging therapies. This study aimed to develop an anti-LIF immunoPET agent for monitoring LIF expression in vivo, improving detection and targeted treatment strategies for glioblastoma. An anti-LIF antibody was conjugated to p-SCN-Bz-DFO and radiolabeled with positron-emitting zirconium-89 (⁸⁹Zr). Target binding properties and stability of the radioimmunoconjugate were assessed by ELISA and size exclusion chromatography. The biodistribution of [⁸⁹Zr]Zr-DFO-anti-LIF was evaluated by PET/CT imaging in an orthotopic glioblastoma mouse model with LIF-positive (GL261N) and LIF-negative (GL261N-CRISPR/LIF) tumors at 24, 48, and 72 hours post-administration. Tumor LIF levels were measured ex vivo by immunohistochemistry. Mass spectrometry determined 2.4 ± 0.3 chelators per antibody molecule. Competitive ELISA demonstrated unaltered affinity post-conjugation. Radiolabeling at a 1 MBq of ⁸⁹Zr per 5 μg of anti-LIF ratio achieved >68% yield, >95% purity, and 0.17 ± 0.03 MBq/μg specific activity. The radioimmunoconjugate remained >90% intact after 72 h in both saline and mouse serum. PET imaging revealed specific accumulation in LIF-positive brain tumors in vivo (6 ± 1.26 % ID/mL at 72 h), which was 2-fold higher than that observed in the GL261N-CRISPR/LIF model. [⁸⁹Zr]Zr-DFO-anti-LIF was successfully synthesized, exhibiting specificity and stability in vitro and in vivo, thus supporting its potential for glioblastoma monitoring, as well as guiding anti-LIF therapies.

Metastatic evolution of malignant tumors following standard anticancer therapies and the emergence of resistant cancer cell populations remain major challenges in oncology. One promising strategy is to develop compounds that selectively target mechanisms of therapeutic resistance. Unlike therapy-sensitive malignant cells, which rely primarily on glycolysis for energy, many chemoresistant cells and cancer stem cells (CSCs) preferentially utilize mitochondrial oxidative phosphorylation (OXPHOS). In this study, we employed a triple-negative breast cancer model to demonstrate that short antimicrobial peptides can significantly suppress the metastatic potential of resistant cancer cells and reduce the formation of CSC-like mammospheres by disrupting mitochondrial respiration. This effect was further enhanced by conjugating the peptides to the mitochondrial-targeting cation triphenylphosphonium (TPP). Mechanistic studies revealed that these compounds induce oxidative stress and mitophagy and suppress mitochondrial translation. Collectively, these findings suggest that TPP-conjugated peptides represent a promising therapeutic strategy for targeting OXPHOS-dependent resistance in aggressive solid tumors.

Pulmonary fibrosis (PF) is a progressive, life-threatening lung disease marked by excessive accumulation of extracellular matrix (ECM), especially collagen, which leads to lung stiffening and impaired respiratory function. Although antifibrotic drugs like nintedanib and pirfenidone can slow disease progression, their clinical benefits remain modest. Hydronidone, a structural analogue of pirfenidone, has shown reduced/no hepatotoxicity, improved tolerability (in humans), and proven antifibrotic effects in liver fibrosis. This study investigated hydronidone’s therapeutic potential in pulmonary fibrosis through both in vitro and in vivo models. In vitro studies utilizing transforming growth factor-β (TGF-β)-induced fibrotic differentiation in LL29 and DHLF cells revealed that hydronidone (62.5, 125, and 250 μM) significantly attenuated the expression of fibrotic markers at both the gene and protein levels, demonstrating greater efficacy compared with pirfenidone (500 μM). In an in vivo manner, a bleomycin (BLM)-induced PF mouse model was employed. BLM administration led to significant physiological and histological alterations, including body weight loss, elevated lung index, alveolar wall thickening, and excessive collagen deposition. Hydronidone treatment significantly attenuated these pathological changes in a dose-dependent manner. Moreover, it improved the lung functional capacity and suppressed BLM-induced upregulation of fibrotic gene and protein expression in lung tissues. Mechanistic studies revealed that hydronidone binds to TGF-β receptor-1 kinase with a better docking score of −7.245 kcal/mol compared to the ligand binding score and inhibits the TGF-β/SMAD (Suppressor of mothers against Decapentaplegic) signaling pathway, similar to pirfenidone. Hydronidone effectively reduces pulmonary fibrotic marker expression and improves lung function at lower doses (25 and 50 mg/kg) than pirfenidone (100 mg/kg) without compromising the safety profile. These findings support its potential as a promising antifibrotic agent and warrant further clinical investigation.

Arachidonic acid (AA) and its metabolites play critical roles in inflammation and immune regulation, modulating the initiation, amplification, and resolution of inflammation. However, their comprehensive quantification remains a challenging endeavor owing to complex metabolic pathways and biological matrix effects. This study introduces a novel metabolomics method involving 5-(diisopropylamino)amylamine (DIAAA) derivatization coupled with ultraperformance liquid chromatography–tandem mass spectrometry to address these issues. The method demonstrated high sensitivity and specificity, with limits of quantification meeting stringent criteria (relative standard deviation <20%; recovery rate, 85–115%, signal-to-noise ratio >10). It effectively quantified 14 key AA metabolites, including hydroxyeicosatetraenoic acids, prostaglandins, and leukotrienes, across a wide linear range (R² > 0.98). The results of intra- and interassay precision tests exhibited low coefficients of variation (≤15%), underscoring the reproducibility of the method. DIAAA derivatization also mitigated matrix variability, improving the accuracy of metabolite detection in serum samples. The hallmark of allergic diseases is a disrupted AA metabolism, where elevated specific metabolites (AA, HETEs, LTB4, and PGD2) show strong diagnostic promise, and a unique metabolite signature in polysensitized patients indicates a link to inflammatory severity. This advanced analytical approach offers significant potential for elucidating the role of AA metabolism in allergic diseases and holds promise for applications in clinical diagnostics and therapeutic monitoring.

Ongoing Alzheimer’s disease (AD) drug development research addresses the need for therapeutic agents that can ameliorate cognitive symptoms and attenuate the course of AD synaptic deficits and neurodegeneration. There is growing interest in evaluating FDA-approved drugs for repurposing as candidate AD therapeutics. Such drugs have the advantage that data are available about their pharmaceutical properties, including doses, pharmacokinetics, pharmacodynamics, biomarkers, metabolism, and safety, to inform the design of clinical drug trials. Importantly, the suitability of such drugs with properties needed for AD requires evaluation. In the early stage of AD, degeneration of the locus coeruleus (LC) brain region results in the reduction of noradrenergic neurons and the loss of the neurotransmitter norepinephrine (NE) that regulates cognition and degeneration. Elevation of extracellular NE through inhibition of the NE transporter (NET) is hypothesized to ameliorate AD deficits. Notably, the NET reuptake inhibitor atomoxetine, an FDA-approved drug for the treatment of attention deficit hyperactivity disorder (ADHD), provides an attractive candidate as an AD therapeutic agent because it may attenuate cognitive decline in AD patients, positively impact AD biomarkers, and reduce neuropathology. The goal of this review is to assess atomoxetine for repurposing in AD based on its ability to improve cognition, regulate NE, impact AD biomarkers, and preserve LC neuronal function, with suitable pharmacokinetics, drug metabolism, and safety based on analysis of clinical and preclinical studies. Evidence for neuroprotective effects of atomoxetine in the early stage of AD at clinically safe doses with suitable pharmaceutical properties supports its candidacy as a repurposed drug for AD therapeutics.