Journal of Pharmaceutical Innovation最新文献

Purpose

The present study aimed to develop a novel and green analytical method for the simultaneous analysis of Chloride, Sodium Citrate, Glycine, Niacinamide, D-Panthenol, and Pyridoxine in ORT LRG Liquid. The developed method was thoroughly validated by ICH Q2 (R1) guidelines to ensure its suitability for accurate and consistent quantitative analysis.

Methods

Analysis was performed using a RP-HPLC method with a C-18 column and a gradient elution technique. The method was optimized under suitable chromatographic conditions. The method was validated for system suitability, specificity, linearity, LOD, LOQ, accuracy, precision, and robustness. The method was also assessed using the greenness (AGREE, MoGAPI) and RAPI tools.

Results

The developed method was found to be in accordance with the ICH Q2 (R1) guidelines. The validation parameters remained within acceptable limits. The method demonstrated good suitability (tailing factor ˂ 2%, theoretical plates ˃ 2000), robustness, specificity without any notable interference, exhibited good linearity (R² ˃ 0.990), acceptable LOD, LOQ values, precision (RSD ˂ 2%), recovery rates (˃ 98%), AGREE (0.71), MoGAPI (79), and acceptable RAPI scores confirming the reliability and greenness of the developed method for Chloride, Sodium Citrate, Glycine, Niacinamide, D-Panthenol, and Pyridoxine.

Conclusion

The developed RP-HPLC method proved to be novel, reliable, precise, robust, and can be effectively applied in future research for the quantification of these compounds simultaneously in feed supplements, veterinary and pharmaceutical formulations containing a complex blend of electrolytes, buffering agents, amino acids, and vitamins.

Graphical Abstract

Introduction

The wood apple Limonia acidissima Linn has long been used in Ayurveda to treat urinary and renal conditions. Despite its wide use in traditional medicine, there is no systematic evidence to support its antiurolithiatic effectiveness. By analyzing the protective effect of Limonia acidissima leaf extract in ethylene glycol (EG)-induced urolithiasis and investigating its underlying mechanisms, this study justifies its traditional use.

Materials and Methods

In this study, the antiurolithiatic effects of the ethanolic leaf extract of Limonia acidissima Linn (ELELA) were investigated by assessing its therapeutic efficacy and urine and serum biochemical indicators in a urolithiatic rat model. Wistar albino rats were categorized into five groups (1–5). Urolithiasis was induced in all groups except the normal control group by administering 0.75% v/v EG in drinking water for 28 days. Treatment groups were administered either the usual medication (cystone 750 mg/kg) or ELELA at dosages of 200 mg/kg and 400 mg/kg, respectively. Urine and serum biochemical indicators were assessed, along with calcium and phosphorus concentrations in kidney homogenates. A histopathological examination of the renal tissue was conducted.

Results

EG-induced urolithiatic rats exhibited significant increases in urinary calcium, phosphate, uric acid, serum creatinine, and blood urea nitrogen (BUN), along with decreased urinary pH and magnesium. ELELA treatment, particularly at a dose of 400 mg/kg, significantly reversed these alterations, restored normal renal function, and reduced renal crystal deposition. Histological examination confirmed the protection of glomeruli and tubules, which was comparable to the standard treatment.

Conclusion

Limonia acidissima Linn demonstrated significant antiurolithiatic and nephroprotective effects, likely through antioxidant, anti-inflammatory, and crystal-inhibiting mechanisms attributed to its phytoconstituents. These findings corroborate its historical applications and suggest its potential as a natural therapeutic agent for urolithiasis.

A stability-indicating RP-HPLC method was developed and validated for the simultaneous estimation of Xanomeline (XAE) and Trospium chloride (TRC) in bulk and pharmaceutical formulations. Chromatographic separation was achieved on a C18 column using a mobile phase of acetonitrile and heptane sulfonic acid buffer (pH 2.5) in a 20:80 (v/v) ratio, with PDA detection at 231 nm. The method demonstrated good system suitability with acceptable resolution, theoretical plate counts (> 2000), and tailing factors. Validation in accordance with ICH guidelines confirmed excellent precision (%RSD < 2), accuracy, robustness, and reproducibility, supporting its suitability for routine quality control and stability studies. The forced degradation study demonstrates that the analyte exhibits acceptable stability under a range of stress conditions, including acid, alkali, peroxide, reduction, thermal, photolytic, and hydrolytic environments. Compared to the control sample (100% assay), degradation varied from minimal to moderate depending on the stress applied. In all stress conditions, peak purity parameters remained within acceptable limits, confirming the stability-indicating capability of the method.

Graphical abstract

Background

Melatonin is an important endogenous hormone widely studied for its therapeutic potential, requiring reliable and sustainable analytical methods for its quantification in biological matrices. Conventional analytical techniques often involve complex procedures and environmentally hazardous solvents, highlighting the need for a simple and green bioanalytical approach.

Objective

The present study aimed to develop and validate a simple, sensitive, and environmentally benign bio-analytical method for the quantitative determination of melatonin in human plasma.

Methods

Melatonin was extracted from human plasma using a liquid-liquid extraction technique and analyzed at 225 nm. The method was validated for linearity, precision, accuracy, recovery, selectivity, ruggedness, and robustness in accordance with bioanalytical validation principles. Environmental sustainability was assessed using Analytical GREEnness (AGREE) and Blue Analytical Greenness Index (BAGI) tools.

Results

The method demonstrated good linearity over a concentration range of 2-10 µg/mL with a correlation coefficient (r²) of 0.997. Intra- and inter-day precision values were within acceptable limits, indicating good repeatability and reproducibility. Accuracy studies showed satisfactory recovery across all quality control levels. The extraction procedure was efficient and consistent, with no endogenous interference observed at the detection wavelength. Ruggedness and robustness studies confirmed the stability of the method under minor variations. AGREE and BAGI assessments indicated a high greenness score.

Conclusion

The developed UV spectrophotometric method is simple, reliable, and environmentally sustainable for the quantification of melatonin in human plasma and can be effectively applied for routine bioanalytical analysis.

Graphical Abstract

Triple-negative breast cancer (TNBC) is the most lethal and metastatic form of breast cancer (BC) that displays elevated levels of epithelial-mesenchymal transition (EMT). Standard chemotherapeutics are unable to reduce the metastatic rate; hence, anti-metastatic agents are instantly required. Zeylenone (ZEY) is an eminent anti-tumour and apoptotic agent that is naturally occurring as cyclohexene oxide, which is extracted from the Uvaria grandiflora leaves. However, the anti-proliferative, anti-metastatic, and apoptotic efficacy of ZEY in TNBC cells remains uncertain. Hence, our investigation proposed to study the antiproliferative, adhesive, EMT markers, and apoptotic activity of ZEY (10, and 15 µM/ml) on human TNBC cells MDA-MB-231 and to assess its latent mechanism. The cell toxicity exploration and cell apoptosis of ZEY action on MDA-MB-231 cells were evaluated by MTT, AO/EB, DAPI, PI, adherence assay, and mRNA expression levels. Data unveiled that ZEY (10 and 15 µM/ml) suppresses cell propagation and adhesion; however, it triggers apoptosis by the heightened mRNA intensities of Bax, caspase, while attenuating cyclin-D1, survivin, Bcl-2, and c-myc, in a dosage-relatedly. It generates a Bax/Bcl-2 ratio disparity, which activates the caspase cascade, Cyt-c, and leads to apoptosis. Furthermore, ZEY (10 and 15 µM/ml) inhibits relative gene levels of EMT markers as well as PI3K/AKT/mTOR pathways. According to MD modelling, ZEY docking investigation reveals that it has a low binding energy and a high binding affinity for the apoptotic and cell growth protein indicators mentioned above, as well as greater stability with Bax and mTOR. Our results emphasise that ZEY is beneficial as a protective remedy for BC. Molecular interaction studies of ZEY with apoptotic and cell proliferative proteins present findings that conclude that ZEY has customised the expression of proteins the breast cancer cells.

Intraoral drug delivery systems offer a promising alternative to conventional oral dosage forms by enabling direct systemic absorption and bypassing first-pass metabolism. In this study, diclofenac sodium (DS)-loaded intraoral films were fabricated using fused deposition modeling (FDM) 3D printing combined with a post-printing soaking-loading technique. The effects of soaking time, film pattern, and drug amount on drug loading capacity and in vitro membrane-controlled diffusion were systematically investigated. The physicochemical properties and content homogeneity of the films were evaluated, while drug-polymer compatibility was assessed using FTIR. In vitro membrane-controlled diffusion studies revealed controlled drug diffusion from all film formulations and demonstrated a significant influence of film pattern on release kinetics. After an 8-hour diffusion period, formulations F4 and F6 exhibited drug diffusion rates of 41.3% and 52.7%, respectively, whereas the DS solution showed a markedly higher diffusion rate of 78.1%. Films fabricated with a striped infill pattern displayed enhanced drug diffusion compared to flat-patterned films, which was attributed to increased surface area and the formation of microchannels within the polymer matrix that facilitated solvent penetration. Importantly, the soaking-loading approach enabled efficient drug incorporation without compromising the structural integrity of the printed films. To the best of our knowledge, this study is the first to demonstrate the combined effect of post-printing soaking-loading-based drug incorporation and film pattern design on drug diffusion behavior in FDM 3D-printed intraoral films. Overall, this strategy provides an innovative, non-invasive, and customizable platform for the development of personalized intraoral drug delivery systems.

Purpose

Pulmonary arterial hypertension (PAH) remains a life-threatening disorder requiring innovative therapies to overcome the limitations of conventional sildenafil (SDF) administration. This study aimed to develop and optimize inhalable SDF-loaded novasomes (SDF-NVS) for efficient pulmonary delivery.

Methods

Using a Box–Behnken design, the effects of stearic acid, cholesterol, and Span 60 concentrations were evaluated on vesicle size (VS), entrapment efficiency (EE%), and cumulative release (CR%).

Results

The optimized formulation exhibited nanosized vesicles (205.36 nm), high EE% (73.16%), and sustained drug release (85.61% over 8 h). Transmission electron microscopy confirmed the spherical morphology of the optimum formulation, while the stability studies over three months showed almost no alteration in the physicochemical properties. Histopathological evaluation of lung tissues showed no inflammation, indicating the safety of repeated intratracheal administration. During pharmacokinetic studies, it was found that SDF-NVS offered increased bioavailability when delivered intratracheally in comparison to oral and intratracheal administration of SDF suspension. The SDF-NVS formulation showed a Cmax and AUC_0−∞ that was 3.2 and 5.1-fold, respectively, greater than the oral suspension. Additionally, it increased the half-life of SDF to 11.52 h, which in turn led to a relative bioavailability of 506%.

Conclusion

These results highlight the safety and efficacy of novasomes as a pulmonary nanoplatform for SDF, with improved bioavailability, sustained release, and enhanced therapeutic outcomes in the management of pulmonary arterial hypertension.

Graphical Abstract

Purpose

This study presents the development of supramolecular nanosystems for the topical administration of rutin (RU), focusing on β-cyclodextrin (Cy) complexes and their inclusion in liposomes (CyL) in a 4:1 molar ratio.

Methods

The supramolecular nanosystems were obtained by the “thin film hydration” method, and characterized in terms of size, morphology, encapsulation efficiency, antioxidant activity and skin safety, by DLS, transmission electron microscopy, UV spectrometry, DPPH radical liberation assay and patch test, respectively. Rutin release kinetics was determined by Franz cell experiments.

Results

The resulting CyL loaded with RU formed stable, homogeneous vesicular dispersions, maintaining color, size, and polydispersity over 30 days, with rutin content consistently above 90%. Antioxidant activity, assessed by FRAP and DPPH assays, was significantly enhanced when RU was encapsulated in Cy and delivered via liposomes. In vitro diffusion studies using Franz cells revealed that CyL formulations provided a controlled release profile for RU, characterized by an initial increase followed by a plateau, suggesting vesicle-mediated release modulation. To improve topical applicability, CyL and Cy formulations were gelled with carbopol or xanthan gum, resulting in pseudoplastic, non-Newtonian gels. Gels containing CyL exhibited reduced spreadability and increased adhesiveness, supporting effective skin application. Patch tests on healthy volunteers confirmed good skin tolerability of all formulations.

Conclusions

These findings demonstrate that the combination of cyclodextrin complexation and liposomal encapsulation, followed by gelation, offers a promising dual strategy to enhance the topical delivery and antioxidant efficacy of rutin.