Biochemical pharmacology最新文献

Diabetic kidney disease (DKD) remains a challenge clinically. Prostaglandins (PGs), many of which activate E-prostanoid receptor-3 (EP3), have been found to increase in dietetic kidneys; however, how this influences diabetic renal injury through EP3 has not been clearly elucidated. In this study, diabetes was induced in mice with global or myeloid cell-selective Ep3 deficiency (Ep3^-/-) and their respective controls by a high-fat diet and streptozocin administration. We showed that myeloid cell-selective Ep3^-/- attenuated renal impairments in diabetic mice similarly as global Ep3^-/-. In accompanying with an increase in PG productions and upregulation of Ep3, increased number of M1 macrophages, inflammation, and multiple forms of regulated cell death (RCD, namely apoptosis, necroptosis, and pyroptosis) were detected in diabetic kidneys. However, all the above-mentioned abnormalities were hardly observed in mice with myeloid cell-selective Ep3^-/-. In vitro experiments further revealed that Ep3^-/- or EP3 antagonism decreased not only macrophage M1-polarization sensitive to Rho kinase inhibition but also the production of PGE₂ in such cells of mice and/or humans. Thus, EP3, whose activation may implicate a self-amplification process, is critically involved in diabetic renal injury via promoting Rho kinase-dependent macrophage M1-polarization to mediate inflammation that may further cause the occurrence of multiple types of RCD, delineating the receptor a promising therapeutic target for diabetic renal injury and diseases with similar mechanisms.

Coronaviruses frequently mutate and cross species, rendering most strain-specific drugs and antibodies less effective and leaving familial-level threats unresolved. The nucleocapsid (N) protein, with highly conserved N- and C-terminal domains, plays a central role in viral replication and immune evasion, making it an ideal target for broad-spectrum antiviral development. We designed DNAzyme targeting conserved N gene sequences from SARS-CoV and multiple SARS-CoV-2 variants, identifying N4 with strong in vitro RNA cleavage activity (K_obs = 0.053 min^-1, 54.56% cleavage rate) and 80% N mRNA knockdown in transfected cells. To enhance its pharmacological properties, N4 was chemically modified with FANA, generating FANA4. Compared with N4, FANA4 exhibited a sixfold increase in serum half-life (8 h to 48 h), a twofold improvement in cleavage efficiency, and > 95% cellular uptake. In N-overexpressing and virus-infected cell models, FANA4 suppressed N protein expression by 99.02% and reduced viral replication by 85%. In vivo, intranasal administration decreased lung viral load by 3-fold without observable toxicity. Transcriptomic profiling revealed ERK/MAPK pathway activation, supporting a dual antiviral mechanism of sequence-specific RNA cleavage and host immunity enhancement. These findings highlight FANA4 as a promising, broad-spectrum nucleic acid therapeutic candidate for combating current and emerging coronaviruses.

Heart failure is a chronic condition with a poor prognosis, and the development of new treatments is an urgent necessity. Extracellular heat shock protein 90 (eHsp90) has been observed to increase in heart failure. However, the pathophysiological role of extracellular Hsp90 (eHsp90) in heart failure development remains unclear. Thus, this study aimed to examine the effects of the anti-Hsp90 antibody 1G6-D7, an eHsp90 inhibitor, on cardiac fibrosis induced by pressure overload. Eight-week-old male C57Bl/6N mice underwent transverse aortic constriction (TAC). Beginning 2 weeks after surgery, the anti-Hsp90 antibody or normal IgG was administered intravenously every 2 weeks. Mice treated with normal IgG developed chronic heart failure with severe myocardial fibrosis 8 weeks after TAC. By contrast, administration of the anti-Hsp90 antibody to the TAC mice partially attenuated myocardial fibrosis and improved cardiac function. The fibronectin level in the myocardial tissue and the interaction between Hsp90 and fibronectin increased in the TAC mice treated with normal IgG. Conversely, these pathophysiological changes were mitigated in the TAC mice treated with the anti-Hsp90 antibody. The results of this study suggest that eHsp90 contributes to cardiac fibrosis by mediating fibronectin in the extracellular space. Furthermore, treatment with the anti-Hsp90 antibody attenuated cardiac fibrosis, which is associated with decreased fibronectin levels. The results of this study indicate eHsp90 as a novel extracellular target molecule for treating heart failure. In addition, our findings also suggested that anti-Hsp90 antibody hold considerable promise as potential pharmaceutical agents for the treatment of heart failure by targeting eHsp90.

Hepatocellular carcinoma (HCC) is a lethal malignancy with limited therapeutic options. Cellular senescence exerts a critical role in tumor progression, but the regulatory mechanisms remain unclear. This study investigates the role of pyruvate dehydrogenase phosphatase 1 (PDP1) in modulating senescence-associated malignant progression in HCC. Our study suggests that PDP1 is upregulated in patients with HCC and is significantly associated with poor prognosis. Functionally, PDP1 induces cellular senescence, activates cyclic adenosine monophosphate (cAMP)/Ca²⁺ signaling, and promotes senescence-associated secretory phenotype (SASP)-driven epithelial-mesenchymal transition (EMT), stemness, and malignant progression. Xenograft models further demonstrate that PDP1 enhances tumor growth in vivo, accompanied by activation of senescence-associated pathways, including p16, p21, cAMP, and Ca²⁺. Adenylyl cyclase 5 (ADCY5), a membrane-associated enzyme responsible for catalyzing ATP into cAMP, is identified as a critical downstream mediator of these effects and serves as a major source of intracellular cAMP production. Mechanistically, PDP1 suppression enhances glycolysis and histone H3 lysine 18 lactylation (H3K18la), a recently identified lactate-derived epigenetic modification, leading to activation of DNA methyltransferase 1 (DNMT1), the primary enzyme maintaining DNA methylation patterns, and subsequent ADCY5 promoter hypermethylation and transcriptional silencing. Importantly, glycolysis inhibition restores senescence and reverses PDP1-driven malignant phenotypes. Collectively, these findings identify that PDP1 drives senescence-associated malignant progression in HCC by linking glycolytic regulation, histone lactylation, and DNA methylation to the control of ADCY5 expression and subsequent cAMP/Ca²⁺ signaling, underscoring its potential as a therapeutic target.

The neuroimmune system is known to be pathologically activated after nerve injury and prolonged exposure to opioids. Whereas previous studies have predominantly focused on the involvement of individual chemokine receptors in this phenomenon, dual CCR2/CCR5 blockade with cenicriviroc represents a novel therapeutic strategy towards greater efficacy in alleviating hypersensitivity. In this study, the expression of opioid (MOR, DOR) and chemokine (CCR2, CCR5) receptors in the spinal cord was assessed by immunohistochemistry after chronic constriction injury (CCI) of the sciatic nerve. Analgesic effects of cenicriviroc were evaluated following intraperitoneal administration of cenicriviroc in behavioral tests (von Frey, cold plate) after single (both sexes) and repeated treatment (alone or with morphine in males). Motor coordination and spontaneous locomotor activity were also tested. Spinal protein levels (p-MOR^S363, p-DOR^S363, CCR2, CCR5, IBA1, GFAP) were analyzed by Western blot. Immunostaining showed that CCR5 and MOR were expressed exclusively by neurons, whereas CCR2 colocalized with neurons, astrocytes, microglia, and/or macrophages, and DOR with neurons and astrocytes. A single intraperitoneal cenicriviroc administration alleviated hypersensitivity in CCI-subjected mice. Unlike morphine, cenicriviroc did not induce tolerance over 16 days of repeated treatment. Moreover, cenicriviroc attenuated nerve injury-induced upregulation of IBA1 at all examined time points and reduced GFAP expression at day 16, which was accompanied by a decrease in CCR2 levels. Cenicriviroc exerts sustained analgesia by simultaneously blocking CCR2 and CCR5 - particularly CCR2 signaling in neurons and glia - which appears to be key to its efficacy. These findings highlight cenicriviroc as a promising, translational candidate for neuropathic pain therapy.

This study aimed to systematically investigate the inhibitory effect and potential molecular mechanism of oridonin derivative U07 on gastric cancer cells. By combining in vitro cell experiments and in vivo animal experiments, techniques including Cell Counting Kit-8 (CCK-8) assay, colony formation assay, Annexin V-FITC/Propidium Iodide (Annexin V/PI) double-staining flow cytometry, Western blot, wound healing assay, Transwell invasion assay, fluorescence staining, surface plasmon resonance (SPR), isothermal titration calorimetry (ITC), and molecular docking were used to analyze the impact of U07 on the biological behavior of gastric cancer cells and related molecular pathways. The results showed that U07 exhibited concentration-dependent cytotoxicity against HGC-27 and MKN-45 gastric cancer cells (with IC₅₀ values of 2.5 μM and 2.6 μM, respectively). It could inhibit cell proliferation and induce apoptosis by activating the caspase-3 pathway, while significantly reducing cell migration and invasion abilities. U07 concentration-dependently increased the levels of reactive oxygen species (ROS) and lipid peroxidation, decreased glutathione (GSH) content, and increased malondialdehyde (MDA) content, thereby triggering ferroptosis; this effect could be blocked by the ferroptosis inhibitor Fer-1. Molecular studies confirmed that U07 could specifically bind to PLK4 kinase (KD = 6.2 μM detected by SPR, Kd = 0.65 μM detected by ITC) and inhibit its activity. PLK4 negatively regulates frroptosis pathway by directly binding to and transcriptionally activating Glutathione Peroxidase 4 (GPX4) / ferritin heavy chain 1 (FTH1). By downregulating PLK4, U07 further reduced the expression of ferroptosis marker proteins GPX4 and FTH1, and promoted lipid peroxidation and ROS production; however, overexpression of PLK4 could reverse these effects. In vivo experiments demonstrated that U07 could inhibit tumor growth in a dose-dependent manner, with the high-dose group (6 mg) showing an anti-tumor effect comparable to that of cisplatin, and it could downregulate the expression of PLK4 and GPX4 in tumor tissues. In conclusion, the oridonin derivative U07 exerts anti-gastric cancer effects by inhibiting cell proliferation, inducing apoptosis, and mediating ferroptosis via PLK4, providing a new candidate drug and therapeutic target for gastric cancer treatment.

Carboxylesterase 2 (CES2), a member of the serine hydrolase superfamily, plays a crucial role in catalyzing the hydrolysis of numerous endogenous and exogenous compounds containing ester bonds. The commonly used clinical drug irinotecan (CPT-11) exerts its anti-tumor effect by being hydrolyzed by CES2 to generate SN-38. Epimedium, a widely used traditional Chinese herb with multiple pharmacological properties, has not yet been characterized for its effects on carboxylesterase 2 (CES2). Our study systematically evaluated the three principal bioactive components of Epimedium for their effects on CES2 activity, revealing that icaritin significantly activated this enzyme. In vitro, Western blot and RT-PCR assays demonstrated that icaritin significantly upregulated CES2 expression at both mRNA and protein levels. Furthermore, icaritin further enhanced CES2 expression by activating the PXR pathway and increased the protein level of P53. Molecular docking simulations demonstrated that the interation energy between CES2 and icaritin was significantly higher than that with cisplatin (a reported CES2 activator), which might suggest that CES2 has a higher affinity for icaritin than cisplatin. In vivo studies confirmed that icaritin increased the hydrolytic activity and protein expression of Ces in mouse liver and intestinal tissues with a concentration-dependent manner. In conclusion, icaritin can enhance the hydrolysis of irinotecan in vitro and in vivo, and this enhancement is related to the activation of CES2 and the increase of CES2 gene and protein expression. These findings have important clinical significance for reducing chemotherapy drug resistance in cancer patients. Abbreviations: CES2, Carboxylesterase 2; CRC, Colorectal cancer; CPT-11, Irinotecan; CYP3A, Cytochrome P450 3A; NR, Nuclear receptor; P53, Tumor protein p53; PPAR-α, Peroxisome proliferator-activated receptor α; PXR, Pregnane X receptor; SN-38, 7-Ethyl-10-hydroxycamptothecin; UGT1A1, UDP-glucuronosyltransferase 1A1.

Ulcerative colitis (UC) is a recurrent inflammatory bowel disease characterized by mucosal inflammation. Recently, the incidence rate of UC increases year by year, and patients diagnosed with UC usually have the poor quality of life. The search for effective treatments of UC remains a crucial research priority. Since traditional Chinese medicine (TCM) is an important treatment for UC, and the components of these TCMs were analyzed. Our data indicated that Mairin was the common core component of TCMs with UC therapeutic effects, including Licorice, Paeoniae Radix Alba and Aucklandiae Radix. Recently, only one study reported that the hydroxamate of Mairin prevented colonic inflammation and fibrosis, and the function and the mechanism of Mairin on UC was still obscure. Dextran sulfate sodium (DSS)-induced UC mice were alleviated after Mairin treatment. Mechanistically, RNA sequencing data indicated that Mairin treatment increased the levels of Irf4 and Cd5l. Molecular docking, drug affinity responsive target stability (DARTS) and immunofluorescence experiments were used to verify that Mairin interacted with EGFR and SRC, promoted IRF4 nuclear import in macrophages. ChIP analysis was verified that IRF4, as a transcription factor, interacted with Cd5l promoter, and Mairin treatment increased the mRNA and protein levels of CD5L. CD5L⁺ macrophages exhibited the high level of M2 phenotype markers, and M2-phenotype macrophages alleviated UC. That was to say, Mairin activated IRF4-CD5L pathway, polarized macrophages into M2-phenotype, and alleviated UC. Our study contributes to the exploration the therapeutic mechanism of Mairin and it also may provide insights for new therapeutic medicine of UC.

ADP-ribosylation is a versatile post-translational modification that governs fundamental processes, including DNA repair, transcription, and stress adaptation. Its homeostasis relies on the dynamic interplay between poly(ADP-ribose) polymerases (PARPs), which assemble mono- or poly-ADP-ribose (PAR) chains on target macromolecules, and ADP-ribosyl hydrolases, which dismantle them. Disruption of this balance leads to the accumulation of toxic PAR and cell death, revealing vulnerabilities that can be therapeutically exploited. PARP inhibitors (PARPis) have revolutionised the treatment of homologous recombination-deficient cancers via synthetic lethality. Yet, emerging resistance limits their long-term efficacy, underscoring the need for novel targets within ADP-ribose signalling. The poly(ADP-ribose) glycohydrolase (PARG), the principal enzyme involved in hydrolysing PAR, has emerged as a compelling candidate: its inhibition amplifies replication stress, drives mitotic catastrophe, and selectively kills cancer cells, particularly those reliant on PAR turnover for survival. Elevated PARG expression correlates with aggressive tumours and poor prognosis, positioning it as both a prognostic biomarker and therapeutic target. This review integrates recent structural and biochemical insights into PARG, highlighting the mechanisms of PAR reversal, regulatory control, and potential synthetic lethal interactions. We also discuss the discovery and development of selective PARG inhibitors, which promise to expand the therapeutic landscape, overcome PARPis resistance, and exploit vulnerabilities in replication-stressed cancers. By bridging mechanistic understanding with translational potential, targeting PARG represents a frontier in precision cancer therapy.

Cancer comprises a diverse group of complex diseases driven by genetic and epigenetic alterations that disrupt cellular signaling, metabolism, and cell death mechanisms. Despite significant advances in therapy, challenges such as tumor heterogeneity, treatment resistance, and escape from regulated cell death continue to impede curative outcomes. Among the various modes of regulated cell death, ferroptosis, an iron-dependent mechanism characterized by excessive lipid peroxidation and oxidative stress, has emerged as a promising therapeutic avenue in oncology. Notably, ferroptosis is intricately linked to iron homeostasis, providing a vulnerability that can be exploited by ferroptosis-targeted strategies in cancer, where iron metabolism is often dysregulated. This review provides a coherent account of the molecular mechanisms governing iron regulation and highlights how its imbalance can trigger ferroptosis. Additionally, we detail the molecular mechanisms of ferroptosis and summarize key regulatory networks, including system x_c-, GPx4, and the FSP1/CoQ10/NAD(P)H axis. Further, the role of natural and synthetic ferroptosis inducers is critically discussed, especially their synergistic potential when combined with chemotherapy, radiotherapy, and immunotherapy. Furthermore, this review explores emerging evidence on the regulation of ferroptosis by non-coding RNAs, hormonal regulation of ferroptosis sensitivity, and nanoparticle-based ferroptosis therapeutic strategies. Finally, the clinical relevance of ferroptosis in cancer therapy is discussed. Overall, this manuscript presents ferroptosis as a promising therapeutic avenue, offering new insights into its integration with existing cancer treatment strategies.