Journal of Pharmaceutical Innovation最新文献

Chronic wounds colonized by methicillin-resistant Staphylococcus aureus (MRSA) prolong hospitalization, heighten amputation risk and accelerate antimicrobial resistance (AMR). To counter this, we exploited the acidic pH (4.5-6.0) of infected tissue and employed ICH Q8/Q9 Quality-by-Design (QbD) to fuse mucoadhesive, cationic chitosan (Cs) with high-surface-area, pH-sensitive graphene oxide (GO), creating a nanocomposite that sustainably releases levofloxacin (LVX) at the wound site, thereby reducing dosing frequency and selection pressure for resistance. This QbD-driven study developed Cs-GO nanocomposite hydrogel for pH-responsive LVX release, optimized via Box-Behnken design (BBD) and response surface methodology (RSM) in Design-Expert^® v13.0. Risk assessment (ICH Q9) identified four critical process parameters (CPPs); Cs: GO ratio (X₁), glutaraldehyde concentration (X₂), stirring speed (X₃), and sonication time (X₄) to refine critical quality attributes (CQAs) like entrapment efficiency and particle uniformity, yielding nanoparticles with 82.31% entrapment, 180 nm size, -29.86 mV zeta potential, and polydispersity index of 0.293 (R² >0.85). These LVX-loaded Cs-GO nanoparticles were subsequently dispersed in an HPMC/Eudragit hydrogel matrix (TGLN4) to produce the final nanocomposite hydrogel dressing. The formulation exhibited pH-dependent release kinetics (85% LVX over 20 h at pH 5.5, mimicking infected wounds), driven by ionic gelation, hydrogen bonding, and glutaraldehyde crosslinking, with anomalous diffusion (Peppas n = 0.65). In vitro assessments demonstrated enhanced mechanical properties (viscosity ≈ 3.4 × 10³ cP) and an effective MRSA inhibition (MIC 0.5 µg/mL). The LVX/GO combination produced rapid bacterial killing consistent with oxidative damage reported for GO-based materials. In vivo studies in a rat MRSA wound model confirmed 90% bacterial reduction and complete re-epithelialization by day 21, with fivefold lower pro-inflammatory cytokines versus controls. ICH stability studies affirmed shelf-life viability (minimal degradation over 6 months). This scalable QbD platform advances manufacturable bionanocomposites for topical antimicrobial therapeutics, reducing dosing frequency and AMR risks while enabling a scalable platform with potential for further manufacturing development.

Mutagenic impurities pose significant safety risks and must be controlled to trace levels in pharmaceuticals. Although methods like HPLC and GC techniques are commonly used, specific analytical methods are required to accurately detect these impurities in drug substances or products. This study focuses on the identification and quantification of 1-(3-bromoprop-1-en-2-yl)-2,4-difluorobenzene (bromo impurity), a potential mutagenic impurity in posaconazole. Classified as a Class-3 impurity under the International Council for Harmonization (ICH) guidelines M7, the bromo impurity requires stringent control due to its potential mutagenic impurity. To address this, a sensitive and selective analytical method using gas chromatography coupled with tandem mass spectrometry (GC-MS/MS) was developed and validated. The GC-MS/MS method employs a thermal gradient elution program, a Rtx-624 (30 m, 0.32 mm, 1.80 μm) consisting of 6% cyanopropylphenyl and 94% dimethylpolysiloxane, and helium as the carrier gas at a flow rate of 2.0 mL/min. The method exhibited excellent sensitivity, with a limit of quantification (LOQ) of 0.3 ppm and a limit of detection (LOD) of 0.1 ppm. Validation parameters, including accuracy (80–120%), precision, robustness, specificity, and solution stability, demonstrated that the method is highly reliable and suitable for routine quality control. This advanced GC-MS/MS method ensures the trace-level detection of mutagenic impurities, offering a robust analytical solution for regulatory compliance and enhancing pharmaceutical safety.

Ciprofloxacin, a widely used oral fluoroquinolone antibiotic, requires precise control over crystallization to ensure optimal downstream processing and final product efficacy. The current study elucidates the application of ultrasound-assisted antisolvent crystallization to produce smaller crystals and enhance yield. Concurrent ultrasonic irradiation and antisolvent introduction via peristaltic pump were employed to modulate the precipitation process aiming to minimize the volume-weighted mean diameter (D[4,3]). Optimized ultrasound parameters established as 140 W ultrasonic power, 80% duty cycle, and 10-minute irradiation time, yielded a significant reduction in mean particle size ( D[4,3] = 2.27 ± 0.02 µm) and an increase in yield (80.21 ± 1.36%) at 1:9 solvent to antisolvent ratio, 100 mL/min antisolvent addition rate, 500 rpm stirring rate and 3 h standing time. In contrast, conventional crystallization conducted under identical conditions but without ultrasound produced larger crystals ( D [4,3] = 2.27 ± 0.02 µm) and lower yield (76.43 ± 1.15%). Even the best-case conventional crystallization at 1:9 solvent to antisolvent ratio, 100 mL/min antisolvent addition rate and 1000 rpm stirring rate resulted a yield of 79.04 ± 0.55% and a D[4,3] = 2.27 ± 0.02 µm. Characterization studies confirmed that ultrasound treatment preserved the chemical structure and crystal morphology of ciprofloxacin. With an ultrasonic energy consumption of approximately 0.019 kWh under the optimized conditions, the study demonstrates the economic feasibility and intensification benefits of ultrasound-assisted crystallization of ciprofloxacin.

Purpose

Around the world, fungal infections are a major concern, and the most commonly used medication to treat them is fluconazole. Posaconazole (PCZ) is a BCS class II drug with low solubility and high permeability. Transfersomes are promising nanocarriers for enhancing drug solubility. This study aimed to create and evaluate transfersomal gels with PCZ incorporated to improve solubility and skin permeation for treating fungal infections.

Methods

Transfersomes were prepared by the thin film hydration method (formulations F1–F6) and incorporated into a carbopol 934 gel base. The compatibility of PCZ with different excipients was examined using DSC, XRD, and FTIR analyses. A characterization study, including vesicle size, PDI, zeta potential, and TEM, was used to analyze the transfersomes. The formulations were then incorporated into a carbopol 934 gel and evaluated for pH, viscosity, and antifungal activity.

Results

Formulations F2 and F5 showed high entrapment efficiencies of 82.1 ± 2.17% and 82.78 ± 2.92%, respectively. F5 exhibited the smallest vesicle size (210 ± 10 nm), good spreadability (8.33 ± 0.21 g·cm·s⁻¹), and sustained release (99.10 ± 0.51%). In antifungal studies, F5 demonstrated the highest inhibition against C. albicans (3.5 ± 0.11 cm), while F2 showed slightly greater activity against A. niger (2.8 ± 0.15 cm), both surpassing the free drug (~ 2.5 ± 0.06 cm).

Conclusion

The present study demonstrated that F5 is a promising approach to enhance the solubility of the drug and increase skin permeability through the deeper skin layers as a topical gel formulation. Effective against fungal infections, PCZ-loaded transfersomal gel can be suited for clinical translation, broad-spectrum applications, combination therapy, commercialization, patient-centered formulations, and nanotoxicology research.

Graphical Abstract

Transfersomal Gel Loaded with Posaconazole for Improved Solubility and Antifungal Potency

Objective

This study was aimed at developing Warfarin composites and preliminarily evaluating them as controlled drug delivery (CDD) systems.

Methods

The Warfarin-β-CD-loaded chitosan nanoparticles (WCS) and Warfarin-β-CD-silica composite (WSi) were synthesized through polymeric nanoparticles and sol-gel processes, respectively. Warfarin-β-CD-loaded chitosan nanoparticles were produced through ionic gelation of chitosan (CS) with tripolyphosphate (TPP) in a specific ratio, followed by loading with the Warfarin-β-CD complex. Warfarin-β-CD-silica composite was synthesized through the sol-gel approach by incorporating the inclusion of Warfarin-β-CD into the silica precursor.

Results

X-ray diffraction analysis revealed a crystalline peak of pure Warfarin at 21.9º of 2θ angle. This peak was shifted to 22.5º in Warfarin-β-CD-CS nanoparticles, indicating the presence of the crystalline nature of Warfarin in this composite. In contrast, no sharp peak was observed in the Warfarin-β-CD-silica composite, suggesting the amorphous nature of the silica-based composite. During FTIR analysis, the characteristic carbon-carbon double bonds (C = C) stretching vibrations corresponding to the phenyl groups 1 and 2 of Warfarin were observed at 1617 cm^− 1 and 1570 cm^− 1 wavenumbers, respectively. The asymmetric stretch of CH₂ group was observed at 2940 cm⁻¹ and an asymmetric bend of CH₃ group revealed at 1451 cm⁻¹. At 1681 cm⁻¹ the stretching of the lactone group (C = O) was indicated. The in vitro drug release revealed that pure Warfarin had a rapid release rate (at pH 1.6, it reached 84.6% in just 28 h, while at pH 7.4, it released around 59.95%) when compared to both composites. In vivo investigations revealed that Warfarin-β-CD-silica composite group showed higher PT (61.2 s at day 3 and 33.9 s at day 9) and INR (4.7 at day 3 and 2.2 at day 9) values compared to the positive control and Warfarin-β-CD-CS groups, suggesting improved Warfarin release control under tested conditions.

Conclusions

Warfarin-β-CD-silica composite exhibited a slow and sustained release of Warfarin, making it a promising candidate for the CDD system for Warfarin.

Graphical Abstract

This study evaluated the wound-healing potential of a Shorea robusta (S. robusta) resin nanogel in streptozotocin-induced diabetic rats. Albumin-stabilized S. robusta nanoparticles (S-NPs) were prepared using an anti-solvent nanoprecipitation method and incorporated into a Carbopol-940 nanogel (S-NP-nanogel) with the particle size of 212 ± 12.87 nm and the ZP to − 21 ± 0.89 mV to enhance stability, dermal penetration, and sustained release. The particles are showed spherical, evenly distributed S-NPs inside a smooth gel matrix, whereas FTIR and PXRD verified robust hydrogen-bonding interactions and semi-crystalline structure. In comparison to diabetic control, topical treatment of the S-NP-nanogel significantly decreased wound area (1.83 ± 0.04 mm², p < 0.001) and accelerated wound contraction (98.70 ± 9.52%). Improved oxidative balance and collagen synthesis were validated by connective tissue measures (HPR, HXA, and HUA) and antioxidant markers (GSH, LPO, and MPO). Histology showed substantial collagen deposition and full epithelialization. The higher anti-inflammatory, wound-regenerative, and antioxidant properties of the S-NP-nanogel suggested that it might be applicable as a natural treatment for diabetic wounds.

Graphical Abstract

Purpose

Tacrolimus, a critical immunosuppressive agent used in autoimmune disorders and organ transplantation, requires rigorous stability assessment to ensure therapeutic efficacy. This study aimed to evaluate the stability of tacrolimus capsules under stress conditions using Response Surface Methodology (RSM), correlating degradation with both chemical and colorimetric changes.

Methods

The experimental design included four independent variables: exposure time (A), temperature (B), relative humidity (C), and hydrogen peroxide concentration (D). Two key responses were measured: the percentage of remaining tacrolimus using HPLC-UV, and the color change (ΔE) between untreated and stress-exposed tacrolimus capsules.

Results

The stability of tacrolimus (% remaining) was significantly affected by all variables (A–D), their interactions (AB, AC), and quadratic terms (A², C²). Color change (ΔE) was primarily driven by time, temperature, their interaction (AB), and the quadratic effect of humidity (C²). Logistic regression confirmed ΔE as a highly accurate predictor of degradation (92.6% correct classification, p = 0.007), with each unit increase in ΔE raising the odds of tacrolimus dropping below 95% stability by 8%.

Conclusion

The study demonstrated that RSM effectively models tacrolimus degradation under stress conditions, enabling precise shelf-life predictions based on time, temperature, humidity, and oxidative exposure. Colorimetric analysis (ΔE) was a rapid, non-destructive compare to HPLC. To minimize degradation, tacrolimus capsules should be stored at controlled temperatures (≤ 25 °C) and low relative humidity (≤ 60% RH), with protection from prolonged heat and oxidizing agents.

Graphical Abstract

A cationic derivative of xanthan gum (Ca-XG) was synthesized through grafting of quaternary amine groups, with a degree of substitution of 0.43 confirmed by ¹H-NMR analysis. The modified polymer displayed amphiphilic behavior, characterized by a critical aggregation concentration of 0.25% (w/v), and demonstrated efficient emulsifying properties. Ca-XG was incorporated into fusidic acid emulgels, where sixteen formulations were evaluated for their physicochemical and functional characteristics. All samples showed homogeneous appearance, with pH values ranging between 5.95± 0.297 to 6.83 ± 0.352, consistent with dermal compatibility. The formulations exhibited good spreadability (32.17–66.91 cm²) and stable drug content (1.96–1.99%). Among them, the optimized formulation provided sustained permeation of fusidic acid over 24 h, with a cumulative permeation of 16.67 mg/cm² and a steady-state flux of 0.724 mg/cm².h. Furthermore, diffusion coefficients (0.015–0.051 cm²/h) confirmed the ability of Ca-XG to control drug transport while ensuring hydration of the topical matrix. Overall, these results demonstrate that Ca-XG acts not only as a bio-based emulsifier but also as a multifunctional excipient, supporting controlled drug.

Graphical Abstract

The potential of siRNA to silence aberrantly expressed genes involved in a variety of diseases makes it a promising therapeutic approach in medical communities. However, poor cell membrane permeability and degradation through endo-lysosomal trapping present major obstacles to its clinical application, necessitating the need for an effective delivery vector. Several peptide sequences, known as cell-penetrating peptides (CPPs), have been reported to transport cell-impermeable cargoes across cellular membranes. In this study, we aimed to design a peptide-siRNA complex for efficient cellular delivery of anti-Smad7 siRNA (siSmad7) as a potential therapeutic approach for colorectal cancer. To achieve this, we carried out a comprehensive screening of peptide databases and identified a cationic peptide (GT) derived from the Oct-6 transcription factor, which had previously demonstrated high cytoplasmic accumulation. The GT-siSmad7 complexes were prepared at different peptide to siRNA molar ratios, and the optimal ratio was selected based on the highest siRNA binding efficiency (75%). Investigating the uptake mechanism by flow cytometry revealed the direct translocation of the peptide-siRNA complex across the cell membrane of HCT-116 cells, resulting in approximately 60% downregulation of Smad7 mRNA and about a 50% decrease in Smad7 protein levels, as determined by real-time PCR and western blot analysis, respectively. Reducing Smad7 levels induced apoptosis in nearly 60% of cancer cells. The GT peptide, derived from an endogenous transcription factor, demonstrated energy-independent uptake, and minimal toxicity, making it a promising and biocompatible delivery system for siRNA.

Graphical Abstract

Purpose

Pirfenidone possesses significant anti-inflammatory and anti-fibrotic properties, making it a promising therapeutic agent for ocular fibrotic conditions. However, its clinical application is limited due to a short half-life (< 19 min) in corneal tissue and poor ocular bioavailability. This study aimed to develop and characterize a nanostructured lipid carrier based in situ gel formulation for enhancing the ocular delivery and therapeutic efficacy of PFD.

Methods

NLCs were prepared via hot melt emulsification followed by probe sonication and incorporated into a gellan gum-based in situ gel system. A central composite design was used for formulation optimization. The optimized nanostructured lipid carrier formulation included Compritol^® 888 ATO (1.64%), Capmul^® MCM (0.32%), Poloxamer 188 (0.5%), and Tween 80 (0.5%). The formulation was characterized for particle size, polydispersity index, zeta potential, entrapment efficiency, in vitro drug release, ex vivo permeation, cytotoxicity, bio adhesion, and biocompatibility using Hen’s egg test using choriollantoic membrane assay. Stability studies were also conducted.

Results

The optimized Pirfenidone loaded nanostructured lipid carrier showed a particle size of 90.15 ± 10.2 nm, PDI of 0.155 ± 0.014, zeta potential of -11.4 ± 1.2 mV, and entrapment efficiency of 90.43 ± 2.14%. In vitro release studies demonstrated sustained drug release (84.39 ± 3.41% over 12 h). Ex vivo corneal and scleral permeation were 82.97 ± 3.01% and 77.01 ± 1.98%, respectively. Biocompatibility, cytotoxicity, and stability assessments confirmed the safety and robustness of the formulation.

Conclusion

The developed nanostructured lipid carrier based in situ gel offers a promising strategy for enhancing the ocular bioavailability and therapeutic potential of Pirfenidone, potentially overcoming the limitations of conventional topical administration.

Graphical Abstract