Chaihu-Shugan-San (CSS), a classic traditional Chinese medicine formula, has demonstrated significant efficacy in treating various gastrointestinal disorders. To explore the therapeutic efficacy of CSS in alleviating chronic atrophic gastritis (CAG), and elucidate the underlying mechanisms of action. High performance liquid chromatography-mass spectrometry was used to identify the main active components of CSS. The therapeutic effects of CSS at doses of 925 mg/kg/day and 1850 mg/kg/day on N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced CAG were evaluated. Network pharmacology and molecular docking were used to predict the potential targets of CSS in CAG. The impact of CSS on the gut microbiota of rats was investigated by 16S rRNA sequencing. The main active components of CSS were lipids and lipid-like molecules, phenylpropanoids and polyketides. In vivo experiments showed that CSS significantly ameliorated MNNG-induced CAG by inhibiting inflammation and apoptosis. The core target of CSS to alleviate CAG were tumor necrosis factor, interleukin (IL)-1β, IL-6, BAX, BCL2, caspase-3/caspase-9, and NFKBIA. Gene Ontology analysis of these core targets revealed their predominant association with the nuclear factor-kappa B (NF-κB) signaling complex and BAX apoptotic complex. Molecular docking demonstrated that six compounds in CSS, including baicalin, licoisoflavone B, licochalcone B, glabrone, glycyrrhiza flavonol A, and marmin exhibited strong binding affinities with NFKBIA. 16S rRNA sequencing indicated that CSS promoted beneficial changes in the colonic microbial community. CSS alleviated CAG by inhibiting NF-κB-mediated inflammation and apoptosis, providing insights into its mechanism of action in protection against CAG.

The global incidence of abdominal aortic aneurysm (AAA) is rising, posing a significant health threat. AAA rupture can lead to life-threatening internal bleeding, yet its underlying pathophysiological mechanisms remain incompletely understood. Furthermore, effective pharmacological interventions to slow or halt AAA progression remain elusive. This study investigates the potential of kaempferol (KA) as a preventive agent for AAA. Using a PPE-induced murine AAA model with preventive KA administration commencing on the day of surgery, we evaluated its effects on AAA formation. Further analysis invivo and invitro assessed KA's impact on macrophage polarization. Network pharmacology was employed to predict key targets, which were subsequently validated through molecular docking, cellular thermal shift assay (CETSA), and rescue experiments. Preventive KA administration significantly attenuated AAA formation, as evidenced by reduced aortic dilation, elastin degradation, and collagen deposition. KA inhibited M1 macrophage polarization (reducing iNOS/CD86) and promoted M2 polarization (increasing Arg1/CD206). Network pharmacology predicted STAT3, STAT1, and TNF as key targets, which was confirmed by molecular docking and CETSA showing direct binding. Importantly, KA inhibited LPS-induced STAT3/STAT1 phosphorylation and TNF expression. Rescue experiments using specific agonists reversed KA's effects on macrophage polarization, demonstrating its action is mediated through the STAT/TNF signaling pathway. Our findings suggest that KA holds promise as a novel preventive strategy for AAA, with its mechanisms of action involving the regulation of macrophage polarization and the STAT/TNF signaling pathway.

Acute lung injury (ALI), a severe inflammatory disease, is intricately associated with the dysregulation of interleukin (IL)-17-mediated inflammatory responses. Terminalia chebula Retz., a traditional medicinal plant, can modulate inflammatory responses. We evaluated the effect of the prophylactic treatment using T. chebula extract (TCE) on lipopolysaccharide (LPS)-induced ALI and its underlying IL-17-linked mechanism. A mouse ALI model was established to assess TCE efficacy by observing histopathological and inflammatory changes. LC-MS identified its invitro and absorbed components, followed by target prediction using integrated network pharmacology, molecular docking, and dynamics simulations, with key findings validated by Western blot analysis. ALI mice, pretreated with TCE, demonstrated notable improvements: lung tissue damage was alleviated and levels of pro-inflammatory cytokines (IL-6, tumor necrosis factor [TNF]-α, IL-17A) were reduced. Immunohistochemistry revealed that TCE markedly suppressed the enhanced alveolar expression of IL-17. Phytochemical analysis identified 55 constituents in TCE, with 15 bioactive compounds (e.g., ellagic acid, chebulinic acid) entering systemic circulation. Multi-method computational analysis confirmed that key compounds, particularly ellagic acid, stably interacted with core targets (e.g., IL-6, CXCL8). The KEGG analysis further highlighted significant enrichment in the IL-17 and NF-κB signaling pathways. These physiological adjustments coincided with the inhibition of the IL-17-MAPK/NF-κB pathway, as evidenced by decreased phosphorylation of p65, p38, and IκBα and downregulation of TLR4, MyD88, iNOS, and COX-2. Altogether, TCE pretreatment effectively alleviates ALI. The modulation of the IL-17-dependent signaling axis may enhance the resolution of inflammation, thereby reducing lung injury. These findings highlight the potential of TCE as a promising therapeutic for ALI via targeting the IL-17-MAPK/NF-κB pathway.

Comment on “Deciphering pathogenic mechanisms of BDCPP exposure in endometrial cancer progression via an integrated approach combining network toxicology, machine learning, and molecular docking”

Osteoarthritis (OA) involves oxidative stress-induced chondrocyte senescence and extracellular matrix (ECM) dysregulation, yet disease-modifying therapies remain elusive. This study investigates the effects of cycloastragenol (CAG), a telomerase-activating triterpenoid from Astragalus membranaceus, on OA progression with a focus on NRF2/NF-κB signaling. In vitro, CAG suppressed oxidative stress-induced senescence in primary rat chondrocytes, evidenced by reduced SA-β-gal positivity, partially restored EdU proliferation, and downregulated senescence related proteins expression. In addition, CAG concurrently attenuated senescence-associated secretory phenotype (SASP) and partially restored ECM homeostasis. Mechanistically, molecular docking analysis suggested a potential interaction between CAG and the Kelch domain of KEAP1. Consistent with this, CAG treatment was associated with NRF2 pathway activation and attenuation of TBHP-induced NF-κB signaling. Importantly, genetic inhibition of NRF2 significantly attenuated the protective effects of CAG, supporting a required role for NRF2 in mediating CAG-induced suppression of oxidative stress and inflammatory signaling. In vivo, intra-articular CAG administration in monosodium iodoacetate (MIA)-induced OA rats reduced cartilage degradation, rescued ECM homeostasis, and enhanced NRF2 activation. Collectively, CAG mitigates the degradation of the extracellular matrix and suppresses the senescence-associated secretory phenotype (SASP) induced by osteoarthritis (OA).

Marine microalgae rich in high-quality proteins are highly favored in the development of bioactive peptides. However, the scarcity of effective preparation methods still limits its application in healthy improvement. This study aimed to search for the effective natural antioxidative peptides from the microalga Porphyridium cruentum (P. cruentum) for protection skin cells against chronic oxidative stress by UPLC-Q-Exactive MS identification coupled to in silico prospection. An antioxidative dipeptide Threonyltyrosine (TY) was identified among 425 peptides. The in vitro antioxidant capacity of TY was slightly higher than that of vitamin C in ABTS radical scavenging assays with an IC50 of (11.37 ± 1.32) ×103 µg/L. The significant protective role of TY against paraquat-induced cytotoxicity in human keratinocyte (HaCaT) cells was observed in cell viability (+ 10.1%, P < 0.001), MDA (- 23.6%, P < 0.0001) levels, and enzyme activities, i.e., superoxide dismutase (SOD, + 17.1%, P < 0.001) and catalase (CAT, + 20.5%, P < 0.001). TY showed a dose-dependent effect (30-100µg/mL) in upregulating the proliferative activity of paraquat-injured HaCaT cells.In addition, TY stable binding to the Kelch domain of Keap1 (ΔG = -5.13kcal/mol) was analyzed via molecular docking and molecular dynamics simulations, which gave some clues on the potential activating Nrf2. Totally, TY exhibited a dual mechanism of free radical scavenging and antioxidant enzymes activation. These findings advanced our understanding of TY's development, properties and provide a foundation for future mechanistic investigations.

Subarachnoid hemorrhage (SAH) is a life-threatening neurological emergency associated with high mortality and poor functional outcomes. Early brain injury (EBI) within the first 72h critically influences prognosis, with excessive microglial activation representing a key contributor to neuroinflammation and secondary neuronal damage. Identifying pharmacological agents capable of modulating microglia-driven inflammatory responses remains an important therapeutic challenge. In this study, we investigated the potential anti-inflammatory mechanisms of pinostrobin (PIN), a natural flavonoid with reported neuroprotective properties. By integrating bioinformatics, network pharmacology, and machine learning approaches (LASSO, random forest, and SVM), MYC and CTSL were identified as candidate targets potentially linking PIN to SAH-related inflammatory pathways. Molecular docking and 50-ns molecular dynamics simulations suggested stable interactions between PIN and these proteins. Furthermore, in vitro evidence from LPS-stimulated BV2 microglial cells demonstrated that PIN reduced the expression of MYC, CTSL, NF-κB p65, and IL-1β, accompanied by attenuation of microglial inflammatory activation. Collectively, these findings suggest that PIN may modulate neuroinflammatory cascades relevant to EBI after SAH, potentially through regulation of the MYC-CTSL axis. This study provides integrative, hypothesis-generating insights into the microglia-targeted anti-inflammatory potential of PIN in the context of SAH.

Molecular docking and chemical composition of commercial essential oils and their toxicity against Spodoptera frugiperda (J. E. Smith, 1797) (Lepidoptera: Noctuidae)

In recent years, cancer has emerged as a significant challenge for healthcare systems, posing challenges to researchers to develop new treatments. Among various cancers, colorectal cancer is a major cause of death worldwide. To develop novel compounds that strongly inhibit colon cancer cells, lonidamine-1,3,4-oxadiazole derivatives 7(a–h) were designed and synthesized. The prepared compounds were characterized by various spectral techniques, including NMR (1H and 13C), mass spectra, and HPLC. Cytotoxicity tests conducted on a colorectal cancer cell line indicated that compound (7d) demonstrated significant antiproliferative effects, achieving the lowest IC50 value of 12.91 ± 1.58 µM, thereby making it the most effective among the compounds tested. This compound induces apoptosis, as evidenced by Hoechst/PI double staining, and mitigates cell migration, demonstrating its antiproliferative and antimigratory capabilities. Molecular docking, dynamics simulations, and DFT studies helped clarify the structure–activity relationship (SAR) and mechanisms of action. ADME and toxicity predictions also supported its drug-like properties. SAR analysis identified key substituents influencing activity, guiding further optimization. Overall, compound (7d) appears to be a promising candidate for colon cancer therapy, though additional studies are necessary to assess its clinical potential. These findings could be used for designing novel cancer therapeutic or preventive LND-derived agents.

Background Dioctyl terephthalate (DOTP), a common plasticizer alternative, has unclear immunotoxic effects on respiratory allergic diseases such as allergic rhinitis (AR), asthma (AS), chronic rhinosinusitis (CRS), and allergic bronchopulmonary aspergillosis (ABPA). Methods We integrated network toxicology, target mining, bioinformatics, and machine learning to explore DOTP’s potential role in allergic airway diseases. Shared targets were analyzed via PPI networks, enrichment analyses, immune profiling, single-cell data, and molecular docking. Key genes were validated in vitro using human airway epithelial cells. Results DOTP showed potential toxicity including carcinogenicity and nephrotoxicity. It shared 136 targets with allergic diseases, mainly involved in immune and apoptotic pathways. EGFR, BCL2, CFTR, SYK, and C3AR1 were identified as core genes. DOTP showed strong binding to these targets and induced cytotoxicity and gene expression changes in epithelial cells. Conclusion DOTP may promote allergic airway diseases via immune-related and multi-target mechanisms. These findings highlight potential health risks of DOTP exposure and warrant further investigation.

Background Carpal tunnel syndrome (CTS) is a common condition in pregnancy, yet reliable tools for identifying women at high risk—particularly those with gestational diabetes mellitus (GDM)—remain lacking. Although GDM shares metabolic features with type 2 diabetes, a recognised CTS risk factor, whether GDM itself causally increases CTS risk and through which molecular pathways has not been established. Methods We used linkage disequilibrium score regression and two-sample Mendelian randomization across FinnGen and UK Biobank to evaluate the genetic correlation and causal effect of GDM on CTS. To identify molecular mediators, we integrated CTS GWAS with whole-blood cis-eQTLs and plasma protein QTLs using summary-data MR and Bayesian colocalization. We then characterised trait specificity through phenome-wide MR and quantified mediation effects through two-step MR. Bulk RNA-seq of CTS tissue, single-cell RNA-seq of placental cell from women with and without GDM, murine histology and immunofluorescence, and molecular docking were used to delineate downstream mechanisms and therapeutic potential. Results GDM and CTS showed significant genetic correlation (rg = 0.219). Genetic liability to GDM causally increased CTS risk across discovery, replication, and female-only models, independent of other metabolic or pregnancy-related traits. Multi-omics integration identified CCS as the only gene supported at both eQTL and pQTL levels and revealed its strongest and most specific causal association with CTS. Mediation MR demonstrated that circulating CCS accounts for a substantial proportion of the GDM–CTS effect. Transcriptomic, single-cell, and animal analyses confirmed a CCS-high, inflamed, and collagen-rich microenvironment in CTS, whereas docking analyses indicated that CCS-centred pathways are pharmacologically tractable. Conclusion GDM exerts a causal effect on CTS, largely mediated through CCS-driven oxidative, immune, and fibrotic pathways. CCS emerges as a promising biomarker for risk stratification and a potential therapeutic target. These findings provide a mechanistic foundation for early CTS surveillance and personalised management in women with GDM, addressing an important unmet clinical need in perinatal care.

Structure based designing of potent anticancer pyrrole-triazole chalcone derivatives: synthesis, molecular docking and <i>in silico</i> ADMET evaluation

Introduction Nicotinic acetylcholine receptors (nAChRs) on immune cells are promising therapeutic targets for the treatment of inflammatory diseases and pain. Both α7 and α9* nAChRs (*denotes the potential presence of other nAChR subunits) have been implicated as mediators of the cholinergic anti-inflammatory system (CAS). This study investigated the binding sites of α7-selective ligands on these receptors and their effects on ATP-dependent release of the pro-inflammatory cytokines interleukin (IL)-1β and IL-18 by human mononuclear phagocytes. Materials and Methods The effects of classical ligands (e.g. ACh, nicotine), unconventional (phosphocholine), putative α7-specific ligands (S24795, PNU-282987 and methyllycaconitine), on the ATP-induced IL-1β release were studied in lipopolysaccharide-primed human monocytic THP-1 cells and THP-1-derived macrophages. Electrophysiological two-electrode voltage-clamp measurements were conducted on Xenopus laevis oocytes expressing human α7, α9 or α9α10 nAChRs. Molecular docking was performed using the crystal structure of the homomeric human α7 receptor (PDB ID: 7EKI) and a modeled pentameric assembly of the homomeric α9 extracellular domain (PDB ID: 6HY7). In addition, the homomeric α10 extracellular domain was generated by homology modeling using 6HY7 as the template. Results In cytokine-release experiments, the nAChR agonists efficiently inhibited ATP-mediated IL-1β release. This inhibitory effect was reversed by specific antagonistic conopeptides [V11L;V16D]ArIB (α7 antagonist) and RgIA4 (α9 and α9α10 antagonist), indicating the involvement of nAChRs containing subunits α7, α9 and/or α10. Electrophysiological measurements suggested an interaction of putative α7-specific ligands with human α9* nAChRs. In molecular docking simulations, all tested ligands showed reasonable binding affinity to homomeric α7, α9 and α10 nAChR models near the C-loop region of the binding pocket. Conclusion Our findings provide a more nuanced framework for interpreting the roles of nAChR subtypes in non-neuronal immune modulation, highlighting the complexity and potential importance of α9* nAChRs in the context of inflammation and innate immunity. The results underscore the importance of considering the nAChR subunit α9 when developing α7-selective ligands for immunomodulation and provides novel insights into the role of α9* nAChRs as potential therapeutic targets for inflammatory diseases and pain.

Para-benzoquinone (pBQ) is of growing concern as an emerging redox-active environmental pollutant due to its ubiquitous presence in smoke and combustion byproducts. Recent reports have highlighted its potential role as a redox-driven mitotoxicant, although the involvement of specific mitochondrial protein targets remains unexplored. Here, we investigated the effects of pBQ on human mitochondrial phenylalanyl-tRNA synthetase (hmtPheRS), an essential enzyme required for mitochondrial protein synthesis and linked to severe neurodevelopmental disorders. Our biophysical analyses revealed that pBQ enhanced the formation of covalently modified higher-ordered structures of hmtPheRS by 75% and induced conformational instability, thereby significantly reducing its aminoacylation activity. NMR spectroscopy and molecular docking analyses further supported interactions between pBQ and residues within the catalytic domain of hmtPheRS, indicating the formation of a protein adduct. In parallel, exposure of HEK293 cells to sublethal concentrations of pBQ (20-40 μM) resulted in altered cellular redox homeostasis. It also impaired mitochondrial membrane potential and respiration, disrupted mitochondrial dynamics, and activated mitophagy. Consistent with the broad reactivity of pBQ and its ability to induce oxidative stress, these findings suggest that hmtPheRS is a vulnerable mitochondrial target whose modification may contribute to mitochondrial dysfunction, together with other redox-dependent pathways. Together, this work highlights mitochondrial aminoacyl-tRNA synthetases as an underexplored class of proteins susceptible to redox-active environmental pollutants.

Species of the genus Stachys (Lamiaceae) are recognized for their ethnobotanical importance and chemical diversity. In this study, the essential oil (EOS) and solvent extracts of the endemic species Stachys sparsipilosa were investigated using integrated GC–MS and LC–ESI–QTOF/MS approaches. GC–MS analysis showed that identified constituents accounted for 94.62% of the total oil, with caryophyllene oxide, kauran-16-ol, and cubebol as major components. Targeted LC–MS analysis quantified prominent phenolic compounds, including chlorogenic acid, rutin, and hesperidin, while untargeted metabolomics tentatively annotated 168 metabolites belonging to phenolics, terpenoids, and other classes. Antioxidant capacity was evaluated using complementary in vitro assays, and enzyme inhibitory activities against α-amylase, α-glucosidase, tyrosinase, acetylcholinesterase, and butyrylcholinesterase were assessed in comparison with standard inhibitors. The extracts demonstrated measurable but generally moderate activities relative to the corresponding positive controls. The essential oil exhibited moderate, non-selective cytotoxic effects at relatively high concentrations, whereas solvent extracts showed limited activity within the tested range. Molecular docking analyses were performed as supportive tools to explore possible enzyme–ligand interactions. Overall, S. sparsipilosa displays a chemically diverse metabolite profile associated with composition-dependent bioactivities, providing a basis for further mechanistic and in vivo studies.

Background/Objectives: Citrus limon leaf essential oil (EO) is traditionally used for its calming and cognitive-enhancing properties. Although the chemical composition of C. limon leaf essential oils (EOs) obtained by means of hydrodistillation (HD) and solvent-free microwave extraction (SFME) has been previously characterized, the influence of the extraction method on their neuroprotective efficacy and dose–response effects remains insufficiently explored. In the present study, EOs obtained by means of HD (CEH) and SFME (CEM) were compared for their behavioral, biochemical, and in silico neuroprotective effects against scopolamine (SCOP)-induced cognitive and anxiety-like impairments in adult zebrafish. Methods: Adult Tübingen zebrafish were exposed to CEH or CEM via immersion at 10, 100, and 150 µL/L for 19 days prior to SCOP challenge (100 µM). Cognitive performance was evaluated using the Y-maze and novel object recognition (NOR) tests, while anxiety-like behavior was assessed using the novel tank test (NTT) and novel approach test (NAT). Brain acetylcholinesterase (AChE) activity and oxidative stress markers were quantified. Molecular docking analyses were conducted to investigate interactions between major EO constituents and AChE and monoamine oxidase A (MAO A). Results: Both CEH and CEM significantly attenuated SCOP-induced memory deficits, improved spontaneous alternation and NOR discrimination, and reduced anxiety-like behaviors. These effects were associated with AChE inhibition and restoration of redox balance. Notably, CEM generally exhibited stronger neurobehavioral and biochemical effects at comparable doses. In silico analyses supported these findings, revealing favorable binding affinities of key EO constituents toward cholinergic and monoaminergic targets. Conclusions: This study demonstrates that the extraction method influences the neuroprotective efficacy of C. limon leaf EOs. While both CEH and CEM exert antioxidant and cholinergic modulatory effects, CEM shows enhanced neuroprotective potential in a zebrafish model of SCOP-induced cognitive impairment, supporting the relevance of extraction-dependent biological profiling in EO-based neurotherapeutic research.

Background Head and neck squamous cell carcinoma (HNSC) is a highly heterogeneous malignancy with poor prognosis and frequent recurrence. Beyond tumor-intrinsic alterations, the immune microenvironment plays a decisive role in tumor initiation and progression. However, the causal contribution of systemic plasma proteins to immune regulation and HNSC susceptibility remains poorly defined. Methods We conducted a multi-sample Mendelian randomization (MR) study integrating large-scale plasma proteomics, immune cell phenotypes, and HNSC. Mediation analyses were performed to identify immune cell phenotypes that potentially mediate protein-HNSC associations. The findings were further supported by immune infiltration analyses, molecular docking and molecular dynamics simulations and validation using clinical HNSC specimens, including single-cell RNA sequencing of collected samples, scTenifoldKnk virtual knockout modeling and immunofluorescence staining/histological assessment of HNSC tissues. Results Among 4907 plasma proteins, MR identified prefoldin subunit 2 (PFDN2) as a protective factor against hypopharyngeal carcinoma, with no evidence of reverse causality. Immune phenotype MR analyses revealed CD64 on monocyte (FCGR1A + monocytes) as the only immune trait causally linked to both PFDN2 and cancer risk. Analysis using multiple deconvolution algorithms demonstrated a consistent negative correlation between PFDN2 expression and monocyte infiltration. Single-cell RNA sequencing revealed predominant PFDN2 expression in epithelial tumor cells, whereas FCGR1A expression was restricted to monocytes. Virtual knockout of PFDN2 selectively activated monocyte-associated inflammatory programs. Molecular docking and dynamics simulations supported a stable protein-protein interaction between PFDN2 and CD64. Tissue analyses further confirmed PFDN2 downregulation and CD64 upregulation in HNSC, correlating with advanced tumor grade and stage. Conclusions Our findings establish PFDN2 as a protective plasma protein that restrains HNSC progression by suppressing CD64 on monocyte-mediated inflammatory immune microenvironments, highlighting the PFDN2-CD64 axis as a potential prognostic biomarker and therapeutic target.

Phosphodiesterase-4B (PDE4B) regulates intracellular cAMP and drives pro-inflammatory cytokine production. Novel small-molecule PDE4B inhibitors with improved isoform selectivity are needed to broaden therapeutic options. We report the discovery and validation of SIP0401, a ChemBridge-derived small molecule prioritized via an integrated in silico pipeline combining pharmacophore modeling, molecular docking, and dynamics-based scoring. SIP0401 was identified through virtual screening and ligand efficiency-guided filtering against the PDE4B catalytic domain. Experimental profiling included PDE-Glo™, HTRF cAMP, and differential scanning fluorimetry (DSF) with recombinant human PDE4B. Isoform preference (PDE4A/C/D) was assessed by PDE-Glo™. Aggregation, solubility, and luciferase interference were evaluated. Cellular assays included cAMP (HTRF) and TNF-α ELISA in THP-1 macrophages, and cytotoxicity (MTT) in BEAS-2B cells. Curve fitting used GraphPad Prism. SIP0401 showed high predicted binding affinity (ΔG = - 8.6kcal/mol) and favorable interaction stability in dynamics. It inhibited PDE4B with IC₅₀ = 282 nM (PDE-Glo) and 338.4 nM (HTRF); rolipram controls gave 91.5 nM and 106.6 nM. DSF showed Tₘ shift + 4.2°C; luciferase interference was ≤ 6%. SIP0401 was selective for PDE4B over PDE4A/C/D (IC₅₀s 1.16-1.97 µM; 5.3-7.0×). In THP-1 cells, SIP0401 increased cAMP (EC₅₀ = 2.52 µM) and inhibited TNF-α (IC₅₀ = 3.24 µM). BEAS-2B viability > 90% up to 50 µM. SIP0401, identified by structure-guided virtual screening, demonstrated moderate biochemical preference for PDE4B over other PDE4 isoforms under the tested conditions. Additionally, the observed cellular activity and favorable colloidal/solubility profiles supports its further optimization and in vivo assessment.

Multitarget-directed ligands offer a promising strategy for overcoming tumor complexity through simultaneous modulation of complementary oncogenic pathways. In this work, a novel (E)-6-(3-(4-methyl-2-thioxo-2,3-dihydrothiazol-5-yl)-3-oxoprop-1-en-1-yl)-2H-chromen-2-one (compound 6) was synthesized and evaluated as a dual inhibitor of tubulin polymerization and tumor-associated carbonic anhydrases (CAs) IX and XII. Compound 6 displayed potent antiproliferative activity, particularly against MDA-MB-231 triple-negative breast cancer cells (IC50 = 0.37 µM), with excellent selectivity toward non-tumorigenic cells. Mechanistic studies demonstrated strong tubulin polymerization inhibition (IC50 = 3.40 ± 0.09 µM) and submicromolar inhibition of CA IX (IC50 = 0.102 ± 0.005 µM) and CA XII (IC50 = 0.213 ± 0.004 µM), accompanied by downregulation of CA-IX and CA-XII protein expression. Cellular investigations revealed pronounced G2/M phase arrest and apoptosis induction via mitochondrial signaling and caspase activation. Anti-angiogenic activity was supported by inhibition of endothelial migration and concentration-dependent suppression of VEGFR-2 (Tyr1175) phosphorylation in HUVEC cells. Human liver microsomal assays indicated measurable metabolic stability, while molecular docking and in silico ADMET predictions supported target engagement and drug-like properties. Collectively, these findings identify compound 6 as a promising multitarget anticancer lead integrating antimitotic, metabolic, and anti-angiogenic mechanisms.

Bisdemethoxycurcumin (BDMC) is a natural curcuminoid with higher solubility and stability than curcumin, yet its potential role in lifespan and healthspan regulation remains largely unexplored. This study aimed to investigate the effects of BDMC on lifespan and healthspan in Caenorhabditis elegans (C. elegans) and elucidate the underlying mechanisms. Among major curcuminoids, BDMC exhibited the most potent lifespan-extending and heat stress-protective effects. Continuous BDMC treatment initiated at the L4 stage significantly extended mean lifespan by 17.7%, with the optimal effect observed at 100 μM, and markedly enhanced heat stress resistance. Notably, mid-life BDMC intervention produced the greatest lifespan extension (24.2%). BDMC also improved healthspan-associated phenotypes, including locomotion and fertility, while reducing senescence-associated β-galactosidase activity and lipid accumulation. Network pharmacology analysis identified epidermal growth factor receptor (EGFR) as a key target, which was further validated by RNA interference, qRT-PCR validation, molecular docking, and molecular dynamics simulations. Collectively, these findings demonstrate that BDMC promotes lifespan and healthspan in C. elegans through modulation of EGFR-related signaling pathways, highlighting its potential as a functional food-derived compound for healthy aging.