A document by Zhengwei Zhang. Click on the document to view its contents.

Objective: Cholangiocarcinoma (CCA) is a highly aggressive malignancy with limited therapeutic options and poor prognosis. Erianin has demonstrated broad-spectrum antitumor activity, yet its role in CCA and tumor immunity remains unclear. Methods: The cytotoxic and growth-inhibitory effects of erianin were evaluated in CCA cell lines and xenograft models. Apoptosis induction, mitochondrial dysfunction, and suppression of focal adhesion kinase (FAK) signaling were examined. The effects of erianin on cellular senescence, senescence-associated secretory phenotype (SASP) factor secretion, and senescent cell clearance were assessed. Pharmacological FAK inhibition was applied to validate the FAK-mediated cell growth and senolytic activity of erianin. The therapeutic efficacy of erianin alone or combined with anti-PD-L1 therapy was evaluated in senescent tumor-bearing mice. Results: Erianin exhibited significant cytotoxicity and induced mitochondrial apoptosis in CCA cells. Rather than inducing senescence, erianin eliminated senescent cells by promoting apoptosis and reduced SASP factor secretion. Erianin effectively inhibited FAK phosphorylation, and FAK blockade further enhanced its antiproliferative and senolytic activity. Moreover, erianin reprogrammed senescence-induced macrophage polarization from an M2- to an M1-like phenotype and increased antitumor immune cell infiltration within the tumor microenvironment. Importantly, erianin synergized with anti-PD-L1 therapy to achieve superior tumor control and prolonged survival in vivo. Conclusions: Erianin exerts potent antitumor effects in CCA by inhibiting FAK-mediated cell growth and senolytic activity, thereby enhancing antitumor immunity and immune checkpoint inhibitor efficacy. These findings identify erianin as a promising senolytic and immunotherapy adjuvant for CCA.

The narrow therapeutic window of reperfusion therapy highlight the urgent need for novel neuroprotective strategies against ischemic stroke, wherein ferroptosis is a key contributor to neuronal death in the ischemic penumbra. This study demonstrates that icaritin (ICT) treatment significantly improved functional outcomes, reduced infarct size,and enhanced neuronal viability in middle cerebral artery occlusion (MCAO) rats and oxygen-glucose deprivation and reoxygenation (OGD/R) PC-12 cells. Transcriptomic and biochemical analyses confirmed that ICT’s neuroprotection was associated with the suppression of ferroptosis. Mechanistically, ICT inhibited pathological mitochondrial permeability transition pore (mPTP) opening, thereby preserving mitochondrial membrane potential and attenuating reactive oxygen species (ROS) production. This action was essential, as the protective effect was mimicked by cyclosporin A (CsA) and abolished by lonidamine (LND). Molecular docking revealed stable binding of ICT to voltage-dependent anion channel (VDAC) and cyclophilin D (CypD.) In conclusion, ICT confers neuroprotection by inhibiting mPTP excessive opening, which in turn blocks the downstream ferroptosis cascade. This work elucidates a detailed mechanism for ICT and validates the “mPTP-ferroptosis” axis as a promising therapeutic target for ischemic stroke.

Background and Purpose Dexamethasone as the first-line treatment for neonatal respiratory distress syndrome (NRDS) has been found to simultaneously promote fetal lung mature and induce offspring neurotoxicity. This model informed study is aim to set the therapeutic window of dexamethasone for NRDS treatment. Experimental Approach Pregnant rats were injected with 0.1 or 0.4 mg/kg dexamethasone to simulate clinically equivalent prenatal-dexamethasone-exposure (PDE). Half rats were anesthetized. Surfactant protein A/B of fetal lung were quantified and pharmacokinetic/pharmacodynamics model was developed to determine the minimal effective concentration. The other half rats delivered naturally and the offspring were underwent open field testing. mRNA-sequencing of fetal hippocampus was performed and concentrations-toxicity assay was conducted in H19-7/IGR-IR cells. These data were integrated with maternal-fetal physiologically-based-pharmacokinetic model to determine the maximum tolerated concentration. Population-pharmacokinetic-model was used to propose optimal clinical dosing regimen. Key Results Pharmacokinetic/pharmacodynamics model well characterized dexamethasone-induced fetal lung maturation, indicating a minimal effective concentration as 1.63 ng/mL. Offspring of PDE rats exhibited significant behavioral alterations. mRNA-sequencing analysis revealed that neurotoxicity may be mediated through the Wnt pathway and renin-angiotensin system, with BDNF and CMA1 identified as potential neurotoxic biomarkers. Cell assays suggested a neurotoxic cut-off concentration of 17 ng/mL. Based on therapeutic window of 1.63-17 ng/mL, a half-dose clinical regimen was proposed as the optimal therapeutic strategy. Conclusion and Implications This model-informed precision dosing strategy offers a rational approach to optimize clinical dexamethasone dosing regimen.

Current treatment of patients with asthma or chronic obstructive pulmonary disease (COPD) predominantly involves the use of inhaled bronchodilators (B2 agonists and muscarinic receptor antagonists)(1) and anti-inflammatory corticosteroids (2). Often these drugs are used in fixed dose combination inhalers, either as dual inhibitors (1) or more recently so called “triple inhalers” (1,3). Whilst in many patients this approach is effective in relieving symptoms and providing maintenance treatment, it is also recognised that many patients remain symptomatic despite such treatment (4). Furthermore, there is ongoing concern about the side effect profile of inhaled corticosteroids, particularly when used in patients with COPD where there is an increased risk of infection (5) and in paediatric patients with asthma (6). Moreover, there are now multiple combinations of bronchodilators and inhaled corticosteroids licenced as medicines and they are delivered in a wide range of inhaler devices which can be bewildering to patients and affect adherence to treatment (1,4).

Background and Purpose Apelin and Elabela are endogenous ligands of the apelin receptor (ApelinR/APJ), a GPCR involved in cardiovascular regulation and body fluid homeostasis. Human contains a conserved disulfide bridge linking the N-terminal domain to extracellular loop 3 (ECL3) via Cys19 and Cys281, forming a structural constraint analogous to a fourth extracellular loop (“ECL4”). In related GPCRs of the chemokine receptor family, this motif plays a critical role in ligand engagement and receptor activation. Experimental Approach The functional role of this disulfide bridge was investigated using site-directed mutagenesis (Cys19Ala, Cys281Ala), plasmon waveguide resonance binding assays, and BRET-based biosensors to monitor Gαi activation and β-arrestin-2 recruitment. The impact on endogenous ligands and antibodies targeting ApelinR was evaluated. Key Results Disruption of the disulfide bridge did not affect receptor surface expression but markedly impaired binding of apelin fragments (pE13F, K17F), leading to reduced Gαi signalling and β-arrestin-2 recruitment. In contrast, binding of the Elabela fragment K22P and the subsequent ApelinR signalling activation were largely preserved. Antagonist antibodies (JN241, JN241-Fc) retained activity, whereas the agonist antibody JN241-9-Fc showed strongly reduced efficacy for the mutated receptors. Conclusion and Implications The N-terminal/ECL3 disulfide bridge is a critical structural determinant for apelin binding and ApelinR activation but is less critical for Elabela signalling, supporting distinct modes of ligand engagement. Antibody-mediated agonism is particularly sensitive to alterations of « ECL4 » without affecting its binding, suggesting that this structural constraint is required for optimal ApelinR activation. Our results further demonstrate that ApelinR shares functional similarities with the chemokine receptors with respect to the role of this « ECL4 » domain.

Background and Purpose: Consumption of cannabis during pregnancy has increased, especially oral administration. Given that cannabis compounds readily cross the placenta, there could be unintended developmental consequences, especially when exposed to the modern obesogenic food environment. We explored the long-term effects of maternal voluntary cannabis consumption on adult offspring’s emotional behavioral and microbiota response to high fat diet (HFD) consumption. Experimental Approach: Pregnant mice voluntarily consumed cannabis extract equivalent to 5 mg/kg/day Δ9-tetrahydrocannabinol from gestational day 1.5 until postnatal day (PD) 10. Male and female offspring (PD49) consumed a HFD or low-fat control diet (LFD) for 12 weeks. We measured a battery of emotional behavioral tasks (elevated plus maze, light dark box, open field test, social interaction), changes in gut microbiota in dams and offspring, and inhibitory synaptic transmission in the lateral orbitofrontal cortex (lOFC). Key Results: Pre- and perinatal cannabis exposure (PPCE) did not influence weight gain of offspring on a HFD. Male offspring receiving HFD had decreased risk-taking behaviours that were exacerbated by PPCE. HFD exposure and PPCE had opposing effects on locomotor activity in male mice. Female mice were protected from PPCE influence on emotional behaviour as well as HFD-induced synaptic depression in the lOFC. Changes in offspring microbiota were associated with PPCE-altered maternal-offspring microbial transmission, particularly from maternal fecal microbiota during late gestation. Conclusion and Implications: PPCE sex-dependently influences HFD-induced anxiety and the gut microbiota of offspring. This raises concerns about differential effects of those exposed to gestational cannabis in the modern food environment.

Background and Purpose: Tyrosine kinase inhibitors (TKIs) targeting BCR-ABL1 have revolutionized the treatment of chronic myeloid leukaemia, but their use is associated with cardiovascular toxicity, recently linked to activation of the innate immune response. Although zebrafish are widely used to study drug mechanisms, they are rarely applied to assess endothelial function. Here, we present a methodology to evaluate TKI-induced endothelial dysfunction in zebrafish larvae. Experimental Approach: Endothelial effects of TKIs were examined in zebrafish larvae by measuring vascular permeability and vasoconstriction in the dorsal aorta. Cardiac toxicity was evaluated through assessment of pericardial oedema and cardiac performance (fractional shortening and fractional area change). Expression of inflammatory and endothelial activation markers was analysed, and the contribution of innate immune cells was tested using genetic ablation of leukocytes (neutrophils and macrophages). Key Results: Ponatinib (third-generation TKI) induced vascular hyperpermeability and vasoconstriction in the dorsal aorta, accompanied by pericardial oedema and reduced cardiac fractional shortening and fractional area change. Nilotinib (second-generation TKI) also increased vascular permeability and caused vasoconstriction and pericardial oedema, but without cardiac dysfunction. Imatinib (first-generation TKI) had minimal effects on vascular tone. However, imatinib and asciminib were not effectively absorbed by static immersion in zebrafish larvae. Ponatinib-induced endothelial effects were associated with increased vcam and icam expression and slightly elevated il-1β and tnfa levels. Genetic ablation of leukocytes did not modify ponatinib-induced vasculotoxicity. Conclusion and Implications: Endothelial profiling in zebrafish larvae provides a quantitative in vivo approach to predict vascular toxicity of anti-cancer drugs. Ponatinib showed the strongest detrimental effects on endothelial function, independent of innate immune cell–driven inflammation.

Background and Purpose: Chronic wounds are a major complication of diabetes mellitus (DM), posing a significant burden on healthcare systems worldwide. Despite advances in the field over the past two decades, the incidence of lower-limb amputations due to diabetes remains high. Persistent hyperglycemia sustains a chronic low-grade inflammation, disrupts hormonal homeostasis, and elevates glucocorticoid (GC) levels both systemically and locally, all of which impair wound healing. The enzymes 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) and steroid 11β-hydroxylase (CYP11B1) are key mediators of local GC reactivation and production, respectively, and are upregulated in diabetic wounds. Experimental Approach: To investigate the therapeutic potential of metyrapone (MET), a dual 11β-HSD1/CYP11B1 inhibitor, as a topical pro-healing agent we used a full-thickness excisional wound model in alloxan-induced diabetic mice. Key Results: Daily topical application of MET (1 mg/wound) for 14 days accelerated wound closure, reduced neutrophilic infiltration, restored myofibroblast differentiation, and enhanced collagen deposition. MET suppressed the expression of several pro-inflammatory genes and production of cytokines, such as IL-1β and IL-6, while reducing local GC production in the skin, without affecting its systemic levels. In vitro, MET enhanced fibroblast migration without altering proliferation. Gene silencing via siRNA revealed that both 11β-HSD1 and CYP11B1 contributed to local GC production. Conclusions and Implications: Collectively, these findings support MET as a promising topical therapeutic strategy to improve wound healing in diabetic patients.

The global burden of obesity continues to rise, demanding effective pharmacological strategies for individuals in whom prevention alone is insufficient. Recent incretin-based and multi-agonist therapies have delivered unprecedented weight loss, demonstrating that the simultaneous modulation of multiple metabolic pathways can optimize body weight control and improve obesity-related comorbidities. Yet the biological heterogeneity of obesity, interindividual variability in treatment response, and uncertainties regarding long-term outcomes highlight the need to explore complementary mechanisms and diversify therapeutic options. Oleoylethanolamide (OEA) is an endogenous lipid mediator synthesized on demand by enterocytes from membrane phospholipids in response to dietary lipid intake. Acting primarily through peroxisome proliferator-activated receptor-α (PPAR-α), endogenous OEA functions as a physiological postprandial satiety signal that promotes fatty acid oxidation and coordinates gut–brain communication. Disruption of this pathway-through genetic or dietary manipulations-impairs metabolic homeostasis, underscoring its regulatory role. Pharmacological administration of OEA in preclinical models reduces food intake without inducing malaise, enhances lipid utilization, and ameliorates diet-induced metabolic dysfunction, while recruiting central circuits governing both homeostatic and reward-driven feeding. Early clinical studies report modest but consistent improvements in body weight and selected cardiometabolic markers, although limited trial size and duration preclude definitive conclusions. Clarifying pharmacokinetics, brain exposure, responder phenotypes, and rational combinations with incretin-based therapies will be essential to define OEA’s place within a more personalized and physiologically grounded anti-obesity therapeutic framework.

Immune checkpoint inhibitor (ICI)– associated myocarditis has emerged as a severe and clinically complex immune-related toxicity that poses significant challenges for therapeutic decision-making in routine cardio-oncological care. High-dose corticosteroids remain the first-line therapy, yet their timing, dosage, and tapering require careful clinical judgment. Early initiation substantially improves outcomes, but steroid resistance or delayed response is common and necessitates escalation to second-line immunosuppression. The heterogeneity of available rescue therapies, including mycophenolate mofetil, abatacept, JAK inhibitors, plasmapheresis, and other targeted agents, creates significant uncertainty, as evidence is largely limited to case reports and small case series. Management of ICI-induced myocarditis requires balancing immunosuppression with cancer control, particularly when considering rechallenge. Prognostic uncertainty and limited long-term data necessitate multidisciplinary, risk-adapted care, while robust prospective evidence and standardized algorithms remain urgently needed. This review summarizes current management strategies for ICI–induced myocarditis, encompassing early high-intensity corticosteroid therapy, treatment of fulminant disease, and the use of second-line immunosuppressive agents. Key clinical challenges, including steroid resistance, preservation of antitumour efficacy, ICI rechallenge, and emerging biomarker-driven and translational approaches, are also addressed to support individualized care.

Background and Purpose: The rising prevalence of metformin non-response in type 2 diabetes mellitus (T2DM) presents a major therapeutic hurdle. While baicalin combined with metformin exerts a synergistic hypoglycemic effect, the underlying mechanism of this combination therapy remains unclear. We identified the key regulatory metabolite of the baicalin-metformin combination via metabolomics, and clarified the molecular mechanism by which this metabolite mediates its synergistic hypoglycemic effect. Experimental Approach: We recruited metformin non-responder (NR) T2DM patients, and a humanized mouse model was established via fecal microbiota transplantation (FMT) to mimic clinical NR phenotype. This model was used to evaluate the hypoglycemic effect of baicalin combined with metformin. Through untargeted metabolomics and targeted lipidomics analyses, ceramide was identified as the key regulatory metabolite. In vitro, baicalin-treated HepG2 cells assessed its effect on serine palmitoyltransferase (SPT), ceramide synthesis rate-limiting enzyme . In vivo SPT inhibitor myriocin treatment validated ceramide’s role in regulating metformin responsiveness. Ultimately, we clarified the underlying molecular mechanism by which ceramide modulates the hypoglycemic effect of metformin via western blotting assays. Key Results: Baicalin restored metformin’s hypoglycemic effect in NR mice, correlating with decreased ceramide levels. Baicalin reduced ceramide production by inhibiting SPT, the rate-limiting enzyme in de novo ceramide synthesis. Myriocin co-administration recapitulated baicalin’s effects, confirming ceramide’s central role in metformin non-responsiveness. Ceramide impaired metformin’s hypoglycemic effect by activating pPP2A Tyr307, which inhibited pAMPK Thr172 activation; baicalin blocked this signaling axis by reducing ceramide levels, restoring AMPK activation and metformin responsiveness. Conclusions and Implications: Baicalin inhibits SPT to reduce ceramide production by targeting de novo ceramide synthesis, alleviates pPP2A/pAMPK signaling axis inhibition, thereby enhancing metformin responsiveness. This study confirms baicalin is a potential adjunctive agent for reversing metformin non-responsiveness, identifying ceramide as a key mediator of baicalin’s action and SPT as its direct target.

Background and Purpose: Vitamin E is well characterized as a lipophilic antioxidant; however, its non-antioxidant functions remain largely undefined. This study was designed to investigate the non-antioxidant mechanisms by which vitamin E regulates programmed cell death and to identify its specific molecular target. Experimental Approach: A high-throughput screen identified vitamin E as an inhibitor of TNF-α/SMAC mimetic-induced cell death. The mechanism of vitamin E was investigated using western blotting, immunoprecipitation, ubiquitination assays, drug affinity responsive target stability (DARTS), cellular thermal shift assay (CETSA), isothermal titration calorimetry (ITC), and molecular docking. Its in vivo efficacy was evaluated in a murine model of DSS-induced ulcerative colitis (UC). Key Results: Vitamin E selectively inhibits TNF-α/SMAC mimetic-induced cell death by directly and competitively binding to the BIR3 domain of cIAP1. This interaction prevents cIAP1 degradation and the subsequent formation of the RIPK1-dependent death complex, an effect that is independent of its canonical antioxidant function. In a murine model of colitis, vitamin E treatment significantly alleviated disease severity by preserving cIAP1. Conclusion and Implications: Our findings unveil a non-canonical function of vitamin E as a direct cIAPs antagonist and regulator of cell death signaling, presenting a mechanistic basis for its potential therapeutic application in inflammatory pathologies driven by aberrant IAP degradation.

Long non-coding RNAs (lncRNAs), is a broad class of non-protein-coding RNAs characterised as new regulators of gene expression at the epigenetic, transcriptional, and post-transcriptional level. Thus, lncRNAs are involved in the regulation of physiological processes and the development of human diseases and cancer by modulating proinflammatory, oncogenic and proangiogenic pathways. Inflammation, cancer and angiogenesis are interlinked through a network of signals and pathways which reinforces itself through feedback loops. Inflammation triggers vessel formation to transfer immune cells and nutrients, and the new vessels then supply more inflammatory cells and factors. Chronic inflammation induces oncogenic transformation and angiogenesis induced by tumour-derived signals support tumour growth and provide the route for distal metastasis. Here we review the implication of lncRNAs in these processes and interactions in the gastrointestinal (GI) tract with a special focus on inflammation and cancer angiogenesis. LncRNAs due to their highly cell-, tissue- and disease-specific expression are attractive targets for the development of novel therapeutics with limited off-target effects and toxicity. We discuss new discoveries which highlight the value of lncRNA targeting as therapeutic applications in GI inflammation and cancer angiogenesis and identify gaps in our current knowledge and understanding. We illustrate advancing approaches, their advantages and challenges for the pharmacological targeting of lncRNAs.

Background and purpose: The neurobiological circuits underlying chronic pain states are not fully elucidated, limiting available treatment options. While the central amygdala (CeA) is a key hub for integrating pain and stress signals, the role of dynorphin-expressing neurons within the CeA remains poorly understood. Experimental approach: We investigated the role of the CeA dynorphin-expressing (CeA dyn) neurons in the affective and sensory components of chronic neuropathic pain induced by chronic constriction injury. Using dynorphin-Cre mice, we selectively manipulated CeA dyn neuronal activity through chemogenetic, optogenetic and pharmacological approaches. Behavior was assessed using conditioned place preference, real-time place preference paradigms, and measuring mechanical withdrawal thresholds. Finally, whole-brain clearing and anatomical tracing were used to identify projection targets of CeA dyn neurons. Key results: Chemogenetic inhibition of CeA dyn neurons produced a conditioned place aversion in mice with chronic neuropathic pain but had no effect in sham animals. Conversely, optogenetic activation of CeA dyn neurons induced place preference in neuropathic pain mice, with no effect in sham controls. Importantly, neither manipulation altered mechanical withdrawal thresholds. Pharmacological blockade of KOR signaling abolished the place preference associated with optogenetic activation of CeA dyn neurons in neuropathic pain mice. Notably, KOR antagonism revealed a place preference in sham animals, suggesting that dynorphin signaling from CeA dyn neurons is aversive in the absence of chronic pain. In contrast, inhibition of the entire CeA reduced mechanical withdrawal thresholds in neuropathic pain without producing place preference. Anatomical tracing revealed that CeA dyn neurons project to multiple brain regions implicated in affective and pain-related behaviors. Conclusions and Implications: These findings indicate that CeA dyn neurons selectively regulate the affective, but not sensory, component of neuropathic pain. In this context, dynorphin release from CeA dyn neurons appears to be beneficial by alleviating the affective dimension of the pain experience.

Background and purpose: Cardiomyocyte (CM) cell cycle exit following neonatal stages positively correlates with epigenetic reprogramming that driving postnatal maturation as defined by increased rigidity of the sarcomere and metabolic reprogramming. However, till date, the epigenetic driver(s) for such process has not been identified. In this study, through a small molecule screening, we defined the histone lysine demethylase, Kdm5b, as a major epigenetic driver that controls the CM cell cycle gene expression in neonatal stage and can be reintroduced in adult CMs to induce cell cycle. Experimental approach and key results: By screening a histone modification small molecule library, we identified AS-8531, a Kdm5b inhibitor, as a potent inhibitor of CM cell cycle. Furthermore, Kdm5b knockdown (KD) using shRNA or CM specific knockout mice in naturally proliferating CMs isolated from postnatal day 1 neonatal mouse hearts (NMCM P1) reduced CM proliferation. While Kdm5b overexpression in adult mouse hearts and human heart slices enhances CM proliferation and augment the induced CM proliferation using other cell cycle factors, indicating that Kdm5b is an epigenetic driver that drive CM genetic program that govern CM cell cycle. Mechanistically, RNAseq and CUT&RUN analyses of Kdm5b KD in NMCM P1 revealed that Kdm5b modulates the expression and the enrichment of H3K4me3 methylation around the regulatory regions of the genes involved in cell cycle. Conclusion: Kdm5b functions as an epigenetic driver that regulates the transcription of key factors controlling CM cell cycle. Overexpression of Kdm5b represents a potential strategy to induce regenerative capacity in adult CMs.

In the 1930s the British Pharmacological Society was founded and quickly began to flourish; Ernest Verney, Henry Dale and J.H. Burn were key players. At the same time, with the rise of national socialism in Germany, many pharmacologists were persecuted for their Jewish ethnicity or for political reasons, resulting in a major influx of highly capable pharmacologists into the UK. Using recently discovered images found in the Department of Pharmacology at the University of Oxford and the detailed diary of Wolfgang Heubner from Hannover Medical School in Germany, we have reconstructed the relationships between a small but significant group of British pharmacologists, German pharmacologists and persecuted pharmacologists. On several occasions, individuals from each of these groups met informally in the UK until just two months prior to WWII. There was also a pharmacology meeting in Berlin in 1938 with prominent British participation but without expatriated German pharmacologists. Most astonishingly, in the UK, the three groups of pharmacologists freely exchanged scientific ideas and planned common projects. Interactions with physiologists also played an important role; there is substantial evidence through photos, diaries and meeting records that there were strong personal bonds between these groups during professional and social gatherings. Within these settings, it appears that politics did not overtly disrupt scientific exchange or socialising. From today’s perspective, it might seem almost unbelievable that genuine friendship between representatives of a democratic country, National Socialist party members and Jewish scientists could exist at that time and yet it did. Sir Henry Dale played a major role as mediator between these groups, also fostering the re-establishment of British-German relations after WWII. This story is a striking example of how long-standing collaborations, shared professional identities and collective goals towards advancing science provide a strong bridge between individuals in the years leading up to WWII, and beyond.

Background and Purpose: Hepatocellular carcinoma (HCC) poses a significant threat to human life due to its high mortality and recurrence rates. Human Golgi complex Glycoprotein 1 (GLG1) serves as a biomarker for various cancers, but its role in HCC remains unreported. This study aimed to investigate the function and mechanism of GLG1 in the malignant progression of HCC. Experimental Approach: GLG1 expression in HCC was analyzed using clinical databases, qRT-PCR, and Western blot. The potential roles of GLG1 in proliferation, migration, and invasion were determined via CCK-8, colony formation, wound healing, and Transwell assays. Downstream pathways were identified by gene enrichment analysis and Western blot. Potential GLG1-binding compounds were predicted using the DGldb database, leading to the synthesis and optimization of YHB-5, which was tested for its anti-tumor activity. Key Results: Knockdown of GLG1 significantly inhibited the proliferation, migration, and invasion of Huh-7 and MHCC97H cells. Mechanistically, GLG1 was found to exert its oncogenic role by regulating the P53/P21 pathway. Furthermore, the novel small molecule compound YHB-5 demonstrated potent anti-tumor activity against multiple HCC cell lines. Conclusion and Implications: Our findings reveal that GLG1 promotes HCC progression via the GLG1/P53/P21 axis. The newly synthesized GLG1-targeting compound, YHB-5, represents a promising candidate drug, highlighting the potential of GLG1 as a therapeutic target for the diagnosis and treatment of hepatocellular carcinoma.