16th International Conference on“From Discovery to Well-Being: A Holistic Vision for Global Health” Organized By: School of Pharmacy, School of Allied Health and Department of Hospital Management, Bhawanipur Global Campus Editors Dr. Soumendra Nath Bandyopadhyay Dr. Swarupananda Mukherjee Dr. Adrija Saha Dr. Uday Sankar Chakrabarty Dr. Kaninika Roy Mr. Sankhajyoti Saha
Objective: The aim of the present study to develop, optimize and characterize ethosomal gel formulation loaded with ashwagandha leaf extract for improved transdermal delivery of Withanolide A for enhanced antidiabetic activity Methods: Ethosomes loaded with Ashwagandha leaf extract were formulated by thin film hydration method and optimized by Box-Behnken design. Ethosomes were evaluated for vesicle size, PDI, Zeta potential and entrapment efficiency. Optimized ethosomes were incorporated into carbopol gel base of different concentration and evaluated for various properties such as appearance, homogeneity, pH, viscosity, and spreadability including Ex-vivo skin permeation study. The antidiabetic activity of optimized gel formulation was assessed in vitro using the α-amylase enzyme inhibition assay and compared with the gel formulation loaded with extract. Results: The optimized ethosomes had vesicle size of 59.338± 0.016 nm with polydispersiblity index of 0.412±0.002 and percentage entrapment efficiency of 92.355± 0.024. The ethosomal gel formulation (F3) containing carbopol of concentration 1.5% was chosen as optimized ethosomal gel formulation based on good appearance, required spreadability and viscosity. In addition, it also showed high permeation and sustained release of withanolide A with flux value of 248.5 (µg/cm²/hr) from ethosomal gel formulation in comparison to gel formulation loaded with ashwagandha leaf extract. Ethosomal gel formulation also showed higher percentage of alpha amylase inhibitory activity of 33.66% in comparison to extract loaded into gel base indicating improved antidiabetic activity in vitro. Conclusion: Incorporation ashwagandha leaf extract into ethosomal gel formulation showed enhancement in the solubility and bioavailability of withanolide A for antidiabetic activity and there by proving that ethosomes are potential as nanocarriers in delivering withanolides for the treatment of Diabetes
Objective: This study evaluated the potential immunotoxicity and safety of a novel hexavalent virus-like particle (VLP)-based rotavirus vaccine candidate by assessing the t-cell-dependent antibody response (TDAR) to keyhole limpet hemocyanin (KLH) in Sprague-Dawley (SD) rats. Methods: SD rats (24 males, 24 females) received three intramuscular vaccine injections at antigen doses of 30, 120, or 600 µg at 14 d intervals, followed by two KLH immunizations (on d 29 and d 35). Safety was monitored via clinical signs, body weight, food consumption, gross pathology, organ weight, and plasma KLH-specific immunoglobulin M (KLH-IgM) and immunoglobulin G (KLH-IgG) levels measured by enzyme-linked immunosorbent assay (ELISA) on d 35, d 42, and d 49. Results: Repeated-dose administration of the "Gam-VLP-rota" vaccine was tolerated, causing no mortality, adverse clinical signs, gross tissue pathology, or systemic organ toxicity. Absolute body weight and food intake remained within physiological norms. Comprehensive gravimetric analysis of internal organs revealed no adverse effects on the metabolic, reproductive, or central nervous systems in either sex. One-way ANOVA followed by Dunnett’s post-hoc test demonstrated that female organ weights remained stable, whereas male rats at the 600 μg dose exhibited a localized enlargement of the thymus (p<0.05). To systematically control type I error inflation across multi-dose longitudinal body weight data, post-hoc pairwise comparisons were subjected to Benjamini–Hochberg FDR adjustments, under which somatic variations did not exceed critical thresholds (adjusted q ≥ 0.3195). However, two-way ANOVA on log-transformed datasets revealed highly significant sex ⨯ treatment interactions for KLH-IgG on d 35 (p = 0.007) and KLH-IgM on d 49 (p = 0.028). Within-sex post-hoc testing with Bonferroni correction revealed a dose-dependent humoral response in females: on d 49, all vaccine groups (30, 120, and 600 µg) significantly exceeded the vehicle control group for KLH-IgM (padj ≤ 0.0125), validated by a monotonic Jonckheere–Terpstra trend test (p = 0.00097). Conclusion: The hexavalent VLP-based vaccine was well-tolerated with no overt immunosuppression or systemic toxicity observed. However, the toxicological significance of the observed thymus enlargement and antibody fluctuations requires further investigation.
Objective: The present study aimed to develop and characterise a pH-responsive dual polymeric nanoparticle system of Tofacitinib citrate using chitosan and Eudragit for controlled drug delivery. Methods: Tofacitinib citrate-loaded nanoparticles were prepared by a single emulsion solvent evaporation method and optimised using a Box–Behnken design under a Quality by Design (QbD) framework. The formulations were evaluated for Particle size, Polydispersity Index (PDI), Zeta Potential, Drug Entrapment Efficiency (DEE), and In vitro drug release. Compatibility studies were performed using FTIR, while surface morphology and crystallinity were assessed by SEM and XRD, respectively. Results: The optimised formulation (F14) exhibited a particle size of 191.9±5.81 nm, PDI of 0.3472±0.01, and zeta potential of+34.66±1.57 mV, indicating good stability and uniformity. The drug entrapment efficiency was found to be 93±4.3%. In vitro release studies demonstrated minimal drug release under acidic conditions (6.05±1.10% at 1 h, pH 1.2), followed by sustained drug release reaching 95.6±2.12% at 24 h in phosphate buffer (pH 6.8). XRD analysis indicated a transition of the drug from crystalline to amorphous form, and SEM images confirmed the formation of spherical nanoparticles. Conclusion: The developed chitosan–Eudragit nanoparticle system demonstrated pH-responsive release, providing an in vitro proof-of-concept for the oral delivery of Tofacitinib citrate. However, further in vivo studies are required to confirm its therapeutic performance.
Objective: To identify novel small-molecule inhibitors of human glyoxalase-I (Glo-I), a zinc-dependent enzyme implicated in cancer-cell detoxification and overexpressed in several cancers, using an integrated structure-based virtual screening and in vitro enzyme-inhibition approach, and to characterize the binding stability of both active hits by molecular dynamics (MD) simulation. Methods: Structure-based virtual screening of the Otava chemical library was performed against the high-resolution human Glo-I crystal structure (PDB: 3W0T, 1.35 Å) using the LibDock and CDocker algorithms in BIOVIA Discovery Studio (DS). Thirteen prioritized hits were evaluated for Glo-I inhibitory activity in vitro against recombinant human Glo-I (rhGlo-I), with myricetin as the positive control, and the binding stability of both active compounds was further examined by 100-ns MD simulations in GROMACS. Results: Of the 13 compounds evaluated, two showed measurable Glo-I inhibitory activity (15% hit rate). Compound D7015500159 showed the highest potency (IC₅₀ = 34.95 µM). Over 100 ns of MD simulation, the D7015500159–Glo-I complex remained stable, with a backbone root-mean-square deviation (RMSD) below 2 Å, favorable solvent-accessible surface area (SASA) values, and up to 10 hydrogen-bond (H-bond) interactions observed in a single frame. In contrast, the D7015870006–Glo-I complex exhibited substantial ligand-RMSD fluctuations (2–8 nm) and did not remain stably bound, consistent with its comparatively weaker inhibitory potency. Conclusion: The two active hits identified here, particularly D7015500159, represent candidate Glo-I inhibitors worth further optimization as anticancer agents, and illustrate that combining structure-based virtual screening with experimental validation is an efficient strategy for discovering inhibitors of tumor-specific metabolic pathways such as the glyoxalase system.
