Acta Pharmacologica Sinica最新文献

Resistance of non-small cell lung cancer (NSCLC) to EGFR tyrosine kinase inhibitors (TKIs) limits the efficacy and leads to disease progression with mechanisms such as activation of autophagy in tumor cells, but the current therapeutic strategies are unable to intervene in this mechanism. Magnolin (Mag), a naturally derived compound, has garnered significant interest due to its potential antitumor properties. Through virtual screening methods, Mag was identified as a compound with potential to regulate molecular pathways closely related to drug resistance mechanisms. In this study, we investigated the ability of Mag to enhance EGFR TKI efficacy in resistant NSCLC. Afatinib-resistant cell line (HCC827AR) was established by continuously exposing HCC827 cells to afatinib (4 µM) for 6 months. Medium containing 4 µM afatinib was refreshed every 48 h. By conducting RNA sequencing (RNA-seq) and exome sequencing on HCC827AR cells, NRG2 was identified as a core-enriched gene. We demonstrated that Mag directly bound to the TYR112 residue of NDRG1, stabilizing its expression and preventing its degradation. This interaction upregulated NDRG1, which in turn promoted its interaction with the E3 ubiquitin ligase HECW1, facilitating the ubiquitination and degradation of NRG2 at lysine 223 (K223). By targeting the NDRG1-NRG2-HECW1 pathway, Mag uniquely inhibited autophagy and restored the sensitivity of HCC827AR cells to EGFR TKIs, thereby reversing resistance. In vivo, the combined treatment with Mag (30 mg· kg^-1 ·d^-1, i.g.) and Afa (10 mg· kg^-1 ·d^-1, i.g.) significantly reduced tumor growth in patient-derived xenografts without inducing major toxicity. This study unravels the intricate role of NDRG1 in modulating NRG2 via HECW1. The results not only illuminate Mag's promising potential as an adjunctive therapy to surmount EGFR TKI resistance, but also underscore the significant therapeutic potential of targeting the NDRG1-NRG2-HECW1 pathway as a novel strategy to reverse EGFR TKI resistance in NSCLC.

Histone deacetylases (HDAC) inhibition represents one of the few validated strategies in epigenetic cancer therapies, demonstrating significant clinical efficacy in T-cell lymphomas and multiple myeloma, yet exhibiting limited efficacy against solid tumors. GCJ-490A is a novel HDAC inhibitor discovered by medicinal chemists in our institute, which exhibits potent in vitro and in vivo anticancer activity. In this study, we investigated the effects of GCJ-490A on the tumor microenvironment and its potential in synergy with PD-1 antibody in anti-tumor therapy. In syngeneic murine models of breast (EMT6) and lung (LL/2) cancers, we demonstrated that GCJ-490A alone and in combination with PD-1 antibody inhibited tumor growth by regulating T cells and tumor-associated macrophages (TAMs). Specifically, GCJ-490A significantly enhanced T-cell proliferation and cytotoxicity, evidenced by the increased expression of Ki67, CD107a and Granzyme B, and modulated TAMs towards a pro-inflammatory M1 phenotype, while reducing the M2 population. In addition, GCJ-490A upregulated PD-1 on T cells and PD-L1 on myeloid-derived suppressor cells (MDSCs) and TAMs, potentially enhancing PD-1 blockade efficacy. However, the anti-tumor efficacy was less pronounced in LL/2 tumors than in EMT6 tumors, which might be related to the increased infiltration of MDSCs in LL/2 tumors. GCJ-490A promoted MDSCs migration into the tumor by promoting the secretion of CXCL7 from LL/2 cells. In conclusion, GCJ-490A exerts its anti-tumor efficacy by reprogramming the tumor immune microenvironment in EMT6 and LL/2 tumor models, which is augmented when combined with anti-PD-1. However, CXCL7-mediated tumor-type-dependent recruitment of MDSCs by GCJ-490A may limit its therapeutic efficacy, and inhibition of the CXCL7/CXCR1/2 pathway might offer new strategies to address this challenge.

Ion channels are transmembrane protein complexes that control ion transport across the membranes and play a pivotal role for maintaining cellular homeostasis as well as in virus-host interactions. As obligate parasites, viruses hijack the host's ion channel network with spatiotemporal precision to drive their life cycle. In this review, we summarize the key function of ion channels in this dynamic interplay. We then delve into the ways by which different ion channel types facilitate discrete stages of viral infection, including entry, genome release, replication, assembly, and release. By examining dynamic changes in ion channel activity during infection, we reveal how viruses manipulate host ion channels to regulate their life cycle. Moreover, the clinical potential of targeting host ion channels as an innovative antiviral strategy is highlighted. The objective of this review is to comprehensively elucidate host ion channel-virus interactions, as well as the potential of existing ion channel modulators as antiviral drugs, laying the theoretical foundation for the development of novel antiviral therapies.

IL-21, as an immune agonist, has demonstrated limited therapeutic efficacy in cancer immunotherapy. To overcome this inherent limitation, we constructed a yeast surface-displayed IL-21 mutant library guided by the structure of the IL-21/IL-21 receptor (IL-21R). Following one round of magnetic bead sorting and three rounds of fluorescence-activated cell sorting, several variants were isolated and evaluated for receptor-binding affinity at the protein level. We identified an IL-21 Variant (IL-21V) featuring four critical mutations (Q19L, Y23N, V69T and K73Q) at the interaction interface with IL-21R. Compared with wild-type IL-21, IL-21V exhibited an approximately 7-fold increase in IL-21R binding affinity and a ~50-fold increase in STAT3 signalling pathway activation. In preclinical in vivo models, IL-21V exhibited broad-spectrum antitumor activity against B16F10 melanoma, MC38, and CT26 colorectal cancer. Notably, IL-21V exhibited significantly stronger antitumor effects compared to wild-type IL-21, even at a low dose of 0.15 mg/kg, highlighting its potential as a promising and effective immunotherapeutic candidate for cancer treatment.

The stimulator of interferon genes (STING) is a crucial pattern recognition receptor that activates innate immunity, particularly in response to pathogen infection and various stimuli. Notably, activation of STING exhibits remarkable potential in enhancing anti-tumor immunity, underscoring the significance of discovering STING small molecule agonists. Recently, zinc pyrithione (ZPT), a marketed antifungal small molecule, has been reported to possess anti-tumor activity through various mechanisms. Our preliminary screening of STING agonists revealed that ZPT could significantly induce STING activation. In this study, we investigated whether ZPT exerted anticancer effects as a small molecule activator of STING. We showed that ZPT bound to the STING protein in vitro with K_D value of 2.72 μM, and ZPT (1-16 μM) dose-dependently activated the STING-TBK1-IRF3 signaling axis in THP-1 cells. In MC38 tumor-bearing wild-type C57BL/6 mice with normal immune systems, administration of ZPT (5, 10, or 20 mg/kg, i.p., every two days for 14 days) dose-dependently inhibited the tumor growth, activated CD45⁺, CD3⁺, and CD8⁺ T cells in both tumors and spleens, and significantly elevated IL-6 secretion in the peripheral blood. These results highlight the potential of ZPT as an immunotherapeutic agent targeting STING.

Inflammatory bowel disease (IBD), including ulcerative colitis (UC) and Crohn's disease (CD), is characterized by limited treatment options and a therapeutic ceiling. Failure to resolve inflammation is the key driver of disease progression. Formyl peptide receptor 2 (FPR2/ALX), a pivotal mediator of inflammation resolution, has emerged as a promising therapeutic target. In this study, we investigated the expression patterns of FPR2 and clinical relevance in myeloid and lymphoid cells of active IBD patients. By analyzing transcriptomic and single-cell RNA-sequencing data from the GEO database, we revealed aberrant expression of FPR2 and its associated genes in colonic mucosa of IBD patients. We found that FPR2/ALX was highly expressed in the colonic mucosa of UC and CD patients compared to non-IBD controls, strongly correlating with alterations in the MAPK pathway and myeloid cell composition. Notably, high mucosal FPR2/ALX levels were associated with poor response to anti-tumor necrosis factor-α (TNF-α) agent infliximab, and were predictive of disease status (AUC = 0.9143). To assess therapeutic potential, we established a dextran sulfate sodium (DSS)-induced colitis model in wild-type and Fpr2-silenced mice. The mice were orally treated with FPR2/ALX modulators Quin-C1 (QC1) and Quin-C7 (QC7) for 7 days. We showed that oral administration of QC1 or QC7 significantly reduced disease active index (DAI) in wild-type mice, whereas the therapeutic effects were markedly impaired in Fpr2-silenced mice. We conclude that FPR2/ALX may serve as a potential biomarker and therapeutic target for IBD.

Synthetic cannabinoids (SC) target the human cannabinoid receptor 1 (hCB₁) and are extensively metabolized, but the metabolite activity on the hCB₁ receptor after a SC intake is largely unknown. In this study we compared the in vitro hCB₁ receptor activity of 26 metabolites of the synthetic cannabinoid receptor agonists (SCRA) JWH-018, AM-2201, THJ-018 and THJ-2201 as a model system for SC metabolite activity to elucidate their structure-activity relationships. The efficacy and potency of metabolites were assessed using an AequoScreen hCB₁ receptor assay in triplicates and 7-8 concentration points (20 µg/mL-9.5 ng/mL) were used to construct dose-response curves and to determine EC₅₀ and E_max. In silico docking and molecular dynamics were performed using a model of the active form of the hCB₁ receptor with all the metabolites. Final poses were simulated to assess stability under physiological conditions. We showed that carboxylic acid metabolites and 2-hydroxyindole biotransformational products were inactive, while 5-hydroxypentyl SCRA metabolites decreased efficacy to <70%, qualifying them as partial agonists. Eighteen metabolites retained >70% efficacy of their parent compound. Metabolite potencies ranged from 13-3500 nM where the most potent were the 4-hydroxypentyl derivatives of THJ-2201 and THJ-018 and the 4-hydroxyindole derivatives of AM-2201 and JWH-018, also known to be prevalent in vivo metabolites. The efficacy data from in silico experiments were correlated with the in vitro results demonstrating a linear trend (R² = 0.9457), significant (P < 0.0001) at the 95% confident interval between the binding energies and efficacies of the compounds investigated. In silico analysis with docking and molecular dynamics simulations showed that active metabolites maintained a minimum of six amino acid interactions involving all substructures. The in silico molecular dynamics simulations revealed that the efficacy and potency seemed to be driven by a complex network of hydrophobic weak amino acid-ligand interactions. Most prevalent were CH-π interactions and π-π stackings. This study demonstrates the clear structure-activity relationships well correlated to the molecular dynamics simulations, suggesting that metabolites, especially the 4-hydroxy pentyl metabolites, may contribute to the overall effect of SCs in vivo.

Amyloid-beta (Aβ) aggregation is the key component of neuritic plaques that drives Alzheimer's disease (AD) progression and cognitive decline. Although synaptic dysfunction strongly correlates with cognitive impairment, its underlying mechanisms remain unclear. Recently, the kynurenine pathway (KP) of tryptophan metabolism has emerged as a key contributor to AD pathology, and xanthurenic acid (XA), a naturally occurring end-product of the KP, has been implicated in neuroprotection. In this study, we investigated the neuroprotective effects of intranasally administered XA in an Aβ-induced AD mouse model. AD-like pathology was induced in mice by intracerebroventricular injection of Aβ_1-42. The mice received daily intranasal instillation of XA (0.5 μg/5 μL per nostril) for 6 weeks. After XA treatment was completed, the cognitive performance was assessed in behavioral tests, then the mice were euthanized, and the brain were collected for molecular and biochemical analyses. We showed that XA treatment significantly improved the cognitive function of AD mice, and reduced AD-related pathological markers such as APP, Aβ and BACE-1 in the cortex, hippocampus and olfactory bulb. XA treatment also attenuated Aβ-induced oxidative stress through upregulation of the Nrf2/HO-1/SOD1 and key enzymatic antioxidants (GSH, GST, CAT, SOD), while concurrently reducing lipid peroxidation. Furthermore, XA treatment preserved synaptic integrity, evidenced by restoring both pre- and postsynaptic markers (SNAP-25, SYP, SNAP-23, PSD-95) and enhancing signaling via the cAMP-PKA-CREB pathway. Notably, XA differentially modulated metabotropic glutamate receptors, decreasing mGluR2 and increasing mGluR3 expression. In vitro experiments were conducted in APPswe/ind-transfected SH-SY5Y neuroblastoma cells. XA (3-100 µM) dose-dependently improved the cell viability while reducing cytotoxicity and apoptosis. Overall, these results demonstrate that XA confers multifaceted neuroprotection by modulating Aβ pathology, oxidative stress, synaptic function, and glutamatergic signaling, suggesting its potential as a novel therapeutic strategy to mitigate cognitive decline and pathological progression in AD.

Selective serotonin reuptake inhibitors (SSRIs) are characterized by delayed therapeutic onset largely due to their reliance on the desensitization of 5-HT_1A autoreceptors (5-HT_1AR_autos) within the dorsal raphe nucleus (DRN). It has been shown that dissociation of serotonin transporter (SERT) and neuronal nitric oxide synthase (nNOS) interaction selectively modulates 5-HT_1AR_autos, thereby facilitating fast-onset antidepressant responses. Targeting the atypical disk large/ZO-1 (PDZ) domain has been implicated in the SERT-nNOS interaction. In this study, we established a drug screening system based on mBRET combined with biological tests to find SERT-nNOS interaction blockers (SNIBs). During screening the compound libraries, 9 top candidates were found to be capable of binding to the PDZ domain of nNOS. We then identified esflurbiprofen as a promising fast-onset antidepressant candidate. Pharmacodynamic studies revealed that esflurbiprofen effectively penetrated the DRN following systemic administration. Esflurbiprofen (10, 20, 40 mg/kg, i.p., once every 4 days) dose-dependently ameliorated depressive-like behaviors in mice subjected to chronic social defeat stress (CSDS) and chronic restraint stress (CRS). In rs-fMRI analysis, we found that esflurbiprofen enhanced the functional connectivity of emotion-related neural networks in CSDS mice. We further demonstrated that esflurbiprofen disrupted the SERT-nNOS complex in the DRN, augmented membrane-associated SERT, and reduced the concentration of 5-HT in the extracellular space of the DRN. This cascade subsequently enhanced serotonergic neuronal firing through the inhibition of negative feedback mediated by 5-HT_1AR_autos, culminating in an augmented release of 5-HT from serotonergic neurons projecting to the prefrontal cortex and hippocampus. These results highlight the potential of esflurbiprofen to induce rapid antidepressant effects by targeting the SERT-nNOS interaction within the DRN.