The development of protein-based systems capable of delivering bioactive compounds remains an important challenge in pharmaceutical formulation science. In this study, β-lactoglobulin (βLG) was investigated as a biocompatible nanocarrier for norjuziphine, 1-[(4-hydroxyphenyl)methyl]-7-methoxy-1,2,3,4-tetrahydroisoquinolin-8-ol. The binding mechanism, protein structural changes, physicochemical characteristics, drug-release behavior, and biological performance of norjuziphine–βLG nanocomplexes were comprehensively characterized. Fluorescence quenching studies demonstrated spontaneous static complex formation with binding constants in the range of 105–106 L mol−1. Spectroscopic, calorimetric, and computational analyses consistently showed ligand-induced conformational remodeling and stabilization of βLG. The resulting nanocomplexes exhibited spherical morphology with a mean hydrodynamic diameter of 190.1 ± 10.34 nm and an encapsulation efficiency of approximately 79.0%. In vitro release studies demonstrated controlled drug release behavior with moderate pH dependence. Nanoencapsulation increased intracellular norjuziphine accumulation by approximately 2.3-fold relative to free norjuziphine and was associated with enhanced antiproliferative activity in human hepatocellular carcinoma (HCC) cells, a reduced IC50 value (15.3 ± 2.33 μM), and significant regulation of Bax, Bcl-2, and Caspase-3 expression. Collectively, these findings establish a mechanistic relationship between hydrophobic interaction-driven complexation, protein conformational remodeling, and delivery performance, highlighting β-lactoglobulin as a promising protein-based carrier for improving the delivery of bioactive compounds.
Aurora kinase A (AURKA) is a key regulator of mitotic progression and is frequently overexpressed in human malignancies, making it an attractive therapeutic target for the development of targeted anticancer agents. The present study aimed to design and synthesize sixteen novel pyrazole derivatives (Pz-01 to Pz-16) as potential AURKA-targeted anticancer agents and to evaluate their anticancer activity, AURKA-binding potential, and SAR using integrated experimental and computational approaches. The synthesized compounds were screened for cytotoxic activity against human breast cancer (MCF-7 and MDA-MB-231) and cervical cancer (HeLa) cell lines. Among the evaluated derivatives, Pz-10 emerged as the most potent analogue, exhibiting significant antiproliferative activity against MCF-7 cells (IC₅₀ = 22.30 ± 2 μM), comparable to the reference drug doxorubicin, while demonstrating moderate activity against MDA-MB-231 cells (IC₅₀ = 27.60 ± 3 μM). Structure-based molecular docking revealed that Pz-10 adopted a favourable binding orientation within the ATP-binding pocket of AURKA, which was further supported by MM-GBSA binding free-energy calculations and favourable predicted pharmacokinetic properties, indicating excellent oral absorption and drug-like characteristics. To validate the stability of the predicted binding mode, 200 ns molecular dynamics simulations were performed, demonstrating persistent protein-ligand interactions, stable hydrogen-bonding networks, and minimal structural fluctuations throughout the simulation trajectory, consistent with the behaviour of the reference inhibitor. SAR analysis identified hydroxyl substitution on the pyrazole scaffold as a critical determinant of antiproliferative activity and target engagement, whereas bulky halogenated and nitro substituents were associated with reduced biological activity. Collectively, these findings identify Pz-10, together with the structurally related Pz-11, as promising pyrazole-based lead molecules for the development of AURKA-targeted anticancer therapeutics and provide valuable structural insights for the future optimization of selective mitotic kinase inhibitors.
Traumatic brain injury (TBI) induces secondary neuroinflammation driven by oxidative stress, inflammasome activation, and immune remodeling, yet specific mechanism-guided pharmacological interventions remain limited. This study established an artificial intelligence (AI)-integrated network pharmacology and multi-omics framework to evaluate whether hydroxytyrosol (HT), an olive-derived natural polyphenol, may regulate TBI-related neuroinflammatory targets centered on the TXNIP/NLRP3 inflammasome axis. Starting from the SMILES structure of HT, potential targets were predicted using PharmMapper, SwissTargetPrediction, and the Similarity Ensemble Approach and were standardized to UniProt identifiers. TBI-associated genes were integrated from GeneCards, DisGeNET, OMIM, and the Therapeutic Target Database. The overlapping target set was analyzed using STRING-based protein-protein interaction (PPI) networks, MCODE, CytoHubba, Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment. Public GEO transcriptomic datasets (GSE123831 and GSE104687) were used for cross-platform expression validation, differential expression analysis, and exploratory CIBERSORT-based immune infiltration estimation. Random forest (RF), multilayer perceptron (MLP), graph convolutional network (GCN), graph attention network (GAT), SHAP/LIME explainability analysis, LASSO inflammatory-risk scoring, and two-sample Mendelian randomization (MR) were further applied for target prioritization, immune phenotype mapping, and genetic association analysis. Seventy-three overlapping HT-TBI targets were identified. PPI and topology analyses prioritized TXNIP, NLRP3, CASP1, MAPK1, and TP53 as key hubs enriched in inflammasome activation, oxidative stress, apoptosis, and NOD-like receptor signaling. TXNIP, NLRP3, and CASP1 were consistently upregulated in both TBI transcriptomic datasets.LM22-based immune deconvolution suggested increased pro-inflammatory immune signatures and a positive TXNIP-M1 macrophage association (r = 0.63, p < 0.001), which should be interpreted as a transcriptome-derived hypothesis rather than validated murine immune-cell proportions. AI-based models consistently ranked TXNIP/NLRP3 as high-contribution features under internal validation, and removal of these targets reduced model performance. A five-gene inflammatory score achieved an internally evaluated AUC of 0.87, while two-sample MR supported positive genetic associations involving TXNIP expression, TBI risk, NLRP3 and IL-1β expression. Collectively, these findings prioritize the TXNIP/NLRP3/CASP1 module as a computationally supported candidate mechanism through which HT may influence oxidative stress-inflammasome-immune coupling in TBI. This study provides an interpretable drug-target-pathway-phenotype framework and identifies TXNIP, NLRP3, and CASP1 as priority nodes for future experimental validation.
Propofol pharmacokinetics in neonates is characterised by pronounced developmental variability, and performance of existing models in preterm neonates remains insufficiently characterized. We systematically evaluated published propofol population pharmacokinetic models suitable for neonates and developed a new literature-supported meta-model. A new dataset was obtained using opportunistic volumetric absorptive microsampling from preterm and term neonates. Neonates (n = 40, median gestational age: 32.7 weeks) receiving 2 mg/kg propofol for procedural sedation were studied. Blood samples (10 µl) were collected to Mitra™ microsampling devices during 24 h after propofol dose coinciding with routine clinical blood sampling. Propofol concentrations were quantitated with HPLC-MS/MS method. Three published propofol population pharmacokinetic models were compared using numerical and graphical tests of goodness-of-fit and predictive performance. Population modelling was conducted using NONMEM® and the $PRIOR subroutine to describe the time-course of whole-blood concentrations of propofol. No published model adequately described propofol concentration-time course in our new dataset. A sensitivity analysis of clearance maturation structures showed that replacing the base model with alternative formulations of postnatal adaptation (discrete step), postmenstrual age-based sigmoidal maturation, or higher allometric exponents for clearance (1.11 and 1.20 instead of 0.75) decreased the predictive performance of the model. A population pharmacokinetic meta-model incorporating prior information from published neonatal models was developed. Clearance maturation was parameterised as the product of a gestational component and an early postnatal adaptation component. All clearance and distribution parameters were scaled with allometric weight scaling. The final model estimated propofol concentration-time course with reasonable precision and predictive performance in preterm infants.
The d-enantiomeric peptide RD2 (named Contraloid or PRI-002 in clinical trials) was developed for the direct disassembly of toxic amyloid-β (Aβ) oligomers, which are the most neurotoxic aggregate species and play a key role in the development and progression of Alzheimer´s disease (AD). PRI-002/RD2 already demonstrated its safety and tolerability in three phase I clinical trials in young healthy volunteers and patients with mild neurocognitive impairment (MCI) or mild dementia due to AD. Results from a phase II clinical trial with PRI-002/RD2 are expected this year, which was designed to demonstrate safety and efficacy of PRI-002/RD2 in patients at an early stage of AD.The objective of this study was to evaluate the effect of age, sex, genotype and food on the pharmacokinetics of intravenous or orally administered RD2 in male and female transgenic APPswe/PS1ΔE9 (APP PS1) (n=62) or wild type (wt) mice (n=169) under fed and fasted conditions. Age and concomitant food intake influenced the pharmacokinetics of RD2. Slightly higher plasma and brain levels were observed in young compared to old wt mice. However, the effect of food on plasma levels depended on the RD2 dose administered. While at lower doses (200 mg/kg) the effect was substantial, plasma levels were 15 times higher in fasted than in non-fasted animals, the effect was only minor at higher RD2 doses (600 mg/kg).The results of this study indicate the important influence of prior food intake on the bioavailability of the compound, which may also apply to patients and is therefore of interest in further clinical development.
