Pakistan Journal of Pharmaceutical Sciences

Abstract: Background: The genus Allium is a chemically diverse source of bioactive secondary metabolites, yet wild populations remain unevenly studied. Wild Allium ampeloprasum L. from Cyprus is underexplored despite evidence that the species contains phenolic, flavonoid and organosulfur constituents with diverse bioactivity. This study provides a locality-specific phytochemical and in-vivo evaluation of its methanolic bulb extract (AAB). Objectives: To characterise AAB and assess its acute toxicity, antinociceptive and local anti-inflammatory activity in Wistar albino rats. Methods: Dried bulb powder (50.0 g) was macerated in methanol, yielding 2.906 g extract (5.812%). Constituents were quantified by LC-MS/MS and screened using SwissADME and ProTox-III. Acute toxicity followed the two-phase Lorke design using 12 rats. Three independent efficacy cohorts (25 rats each; n = 5/group) received distilled water, AAB at 125, 250 or 500 mg/kg, or piroxicam 20 mg/kg. Antinociception was assessed by writhing, hot-plate and formalin tests and local anti-inflammatory activity by formalin-induced paw oedema. Biochemical, haematological, ESR and CRP measurements were performed after the final 5-h paw oedema assessment. Results: Thirteen constituents were quantified, with caffeic acid and trans-ferulic acid showing the highest relative abundances. No mortality occurred up to 5000 mg/kg, although transient clinical signs were observed. AAB reduced writhing by 45.8%, 69.5% and 91.5% at 125, 250 and 500 mg/kg, respectively (adjusted p = 0.0035, p < 0.0001 and p < 0.0001). Hot-plate latency increased significantly only at 500 mg/kg (adjusted p = 0.0067). Paw licking decreased at all concentrations in both formalin phases (adjusted p < 0.0001), with late-phase inhibition of 85.71-95.71%. Paw oedema inhibition at 5-h was 66.67-71.11%. Post-oedema blood analyses showed selected biochemical changes without a consistent concentration-related pattern. Conclusion: AAB demonstrated antinociceptive and local anti-inflammatory activity in rat models. Absence of mortality does not establish safety; further standardisation, mechanistic investigation and longer-duration toxicity studies are required.

Abstract: Pharmaceutical manufacturing remains constrained by legacy paradigms that collapse engineered systems into static legal descriptors and struggle to integrate real‑world evidence. Geopolitical instability, supply‑chain fragility, climate‑driven disruption, declining institutional trust and the accelerating influence of generative artificial intelligence expose that constraint as an epistemic ceiling. This Perspective outlines a professional path for leapfrogging toward a patient‑centric Pharma 5.0 paradigm grounded in ontological honesty and epistemic humility. The argument traces this ceiling across three layers—the individual practitioner, the quality system and the broader ecosystem—and places Deming’s System of Profound Knowledge, Kegan’s orders of consciousness and McGilchrist’s hemispheric account alongside the classical sequence of the nafs and a heart‑centered posture of practice (Dil Se).  Leapfrogging is a knowledge strategy before it is a technology strategy. Recent United States hearings and independent quality‑scoring efforts reveal an ontology capable of detecting divergence yet unable to revise certified reality. Pakistan and the broader Global South live under the same ceiling, shaped by high generic volumes and thin margins. Thailand’s May 2025 registration of a domestically developed trastuzumab biosimilar demonstrates capability‑first sequencing; a subsequent United States draft guidance issued on 29 October 2025—proposing that comparative efficacy studies may be unnecessary when analytical comparability is high—confirms the direction of that sequence rather than authorizing it after the fact. Preparing for Pharma 5.0 requires unlearning certain validated habits, cultivating sovereignty of judgment before sovereign infrastructure and building quality systems capable of revising their own descriptions.

Abstract: Background: Liver fibrosis is a chronic wound healing process of liver injury and gut–liver axis dysregulation has been recently identified as an important contributor to liver fibrosis progression. Quercetin has been reported to have anti-inflammatory activity and microbiota-modulating effects. However, the potential targets of quercetin remain unclear, as does the relationship between quercetin-associated targets and changes in the gut microbiota in liver fibrosis. Objectives: The purpose of this study was to use network pharmacology to predict potential targets of quercetin associated with liver fibrosis and to explore changes in gut microbiota following treatment with quercetin using a CCl₄-induced rat model. Methods: Eighteen male Sprague–Dawley rats were randomized into CON group, MOD group and QR group (n = 6 per group). Liver fibrosis was induced by weekly subcutaneous injection of carbon tetrachloride (CCl₄) for eight weeks. Oral gavage of quercetin (50 mg/kg/day) was performed in QR group. Histopathological changes were assessed by hematoxylin–eosin staining, Masson staining and Ishak scoring. Potential targets of quercetin were screened using network pharmacology, followed by protein–protein interaction (PPI) network construction and Gene Ontology (GO)/Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. Cecal microbiota composition was determined by 16S rRNA gene sequencing. Parametric or non-parametric tests were used accordingly, including one-way ANOVA or Kruskal–Wallis test. Results: Histopathology revealed significant liver injury and fibrosis in the MOD group relative to the CON group. The QR group showed lower Ishak fibrosis scores and milder histopathological features compared with the MOD group. Network pharmacology analysis identified UBC, UBE2I, RBX1, PSMB7 and UBA1 as hub targets, with KEGG pathways of interest including the TNF signaling pathway, p53 signaling pathway, Toll-like receptor signaling pathway and ubiquitin-mediated proteolysis. Alpha diversity of the cecal microbiota differed significantly among groups, whereas beta diversity did not. Relative abundance of Proteobacteria decreased and Firmicutes increased in the QR group relative to the MOD group. Taxa enrichment analysis identified enrichment of Muribaculaceae and Ligilactobacillus in the QR group. Conclusion: Histopathology and microbiome analyses indicated that quercetin intervention was associated with altered liver histopathological features and shifts in gut microbiota composition in rats with CCl₄-induced liver fibrosis. Network pharmacology predicted several hub targets and pathways potentially linked to these observations. As the present analyses are purely correlational and lack mechanistic validation at the molecular level, no causal relationships are implied between the predicted targets/pathways and the observed histological or microbiome changes. These findings should be regarded as hypothesis-generating and warrant further validation studies.

Abstract: Background: Gallic acid is a natural phenolic compound with excellent antioxidant, antimicrobial and anti-inflammatory activity and good skin penetration. Gallic acid demonstrates both stability and solubility problems in topical formulations. A nanoemulgel combines an emulsion for enhanced solubility with a gel base for improved physical stability. Objectives: The main objective of this study is to characterize and assess the stability of a gallic acid-containing nanoemulgel. Methods: Various gallic acid nanoemulsions (GNE) were prepared using optimized ratios of oil, surfactants, gallic acid and water and evaluated for stability. Finally, stable GNE-19 was characterized for particle size and converted to nanoemulgel (GNEG-19). Nanoemulgel was subjected to short-term and accelerated stability studies and characterized for pH, viscosity, spreadability, occlusion effect, FTIR, in-vitro drug release and in-vitro pharmacological activities. Results: The results showed that formulation GNEG-19 showed optimum thermodynamic stability, having no phase separation and color change. The droplet size of freshly prepared GNE and after 28 days was 175.43nm and 183.23nm, respectively. The pH was slightly acidic but still within the human skin range (3.9–4.5). No interaction was found between the gallic acid and other components of nanoemulgel following FTIR analysis. The viscosity of freshly prepared GNEG-19 was 2127.1 cP and after 90 days, it was 2102.3 cP, which showed its stability. The percent drug release of gallic acid from nanoemulgel after 24 hrs was 93.12%. GNEG-19 also showed effective occlusion properties when compared with gel base and petroleum jelly. In-vitro pharmacological activities showed that GNEG-19 had antioxidant activity of 63.77% and a zone of inhibition (ZOI) of 11.4 mm compared to pure gallic acid (9.5 mm). Conclusion: In conclusion, nanoemulgel containing gallic acid showed good physical and chemical stability and was pharmacologically active.

Abstract: Background: HPV-associated cervical cancer remains a leading cause of cancer-related death in women worldwide. The role of bone marrow mesenchymal stem cells (BMSCs) and miR-129 in tumor progression remains unclear. Objectives: To investigate miR-129 expression and its clinical significance in HPV-associated cervical cancer and to explore whether BMSCs regulate malignant phenotypes through miR-129. Methods: miR-129 expression was measured by qRT-PCR in 77 cervical cancer and 30 normal cervical tissues. Associations with clinicopathological features and overall survival (OS) were analyzed. Diagnostic value was assessed by ROC curve. Rat BMSCs were co-cultured with SiHa and HeLa cells. Following miR-129 overexpression or knockdown, cell proliferation (CCK-8, colony formation), invasion (Transwell) and expression of E-cadherin, N-cadherin and MRP1 (Western blot) were examined. Results: miR-129 expression was significantly higher in cervical cancer tissues than in normal controls (3.24±0.56 vs. 1.00±0.32, P<0.05). High miR-129 expression correlated with tumor size ≥3 cm (P=0.009), FIGO stage III-IV (P=0.0034) and lymph node metastasis (P=0.038). High expression was associated with shorter OS (median: 28.4 vs. 46.2 months; HR=2.944, 95%CI:1.484-5.842, P=0.002). The AUC was 0.788 (95%CI:0.693-0.884), with a cut-off of 1.85 (sensitivity:76.9%, specificity:70.4%). BMSC co-culture upregulated miR-129 expression (SiHa: 3.52±0.42 vs. 1.00±0.08, P<0.01). miR-129 overexpression promoted proliferation, colony formation and invasion, while knockdown suppressed these phenotypes. Overexpression of miR-129 reduced E-cadherin (0.42±0.05 vs. 1.00±0.08, P<0.01) and increased N-cadherin (2.85±0.22 vs. 1.00±0.10, P<0.01) and MRP1 (2.46±0.18 vs. 1.00±0.09, P<0.01). Knockdown of miR-129 partially reversed BMSC-induced changes in these proteins. Conclusion: miR-129 is significantly upregulated in HPV-associated cervical cancer and shows potential as a diagnostic and prognostic biomarker. BMSCs upregulate miR-129 in cervical cancer cells and miR-129 modulates proliferation, invasion, EMT marker expression and MRP1 expression. These findings suggest that the BMSCs/miR-129 axis may contribute to cervical cancer progression, but further in-vivo and mechanistic studies are needed.