Journal of Applied Pharmaceutical Science

Fungal keratitis is a sight-threatening corneal infection affecting over a million people annually. Natamycin, a polyene extracted from Streptomyces natalensis, remains the only FDA-approved drug for the treatment of fungal keratitis. The conventional treatment involves a 5% ophthalmic natamycin solution, administered at a high frequency during the initial treatment phase and subsequently tapered based on the observed clinical response. Although it demonstrates a broad spectrum of activity, its efficacy is hindered by limited corneal penetration due to its high molecular weight and low aqueous solubility.

This review consolidates current knowledge on fungal keratitis, including prevalence, risk factors, and etiology. It provides a comprehensive evaluation of natamycin, covering its pharmacology, mechanism of action, antifungal spectrum, and clinical applications while addressing the limitations of conventional natamycin therapy and ocular barriers that restrict drug delivery. Furthermore, the review highlights recent advances in both nanotechnology-based and non-nanotechnology-based natamycin delivery strategies to enhance corneal bioavailability.

Despite promising preclinical outcomes in terms of bioavailability and retention, no alternative natamycin formulations have yet progressed to clinical trials or commercialization. Further efforts are required to advance these delivery systems into clinical trials and realize their potential to improve outcomes in fungal keratitis.

The Red Sea is a promising, underexplored habitat for the discovery of new bioactive marine natural products. This review provides a comprehensive survey of sesquiterpenes isolated from Red Sea organisms between 2015 and 2025. It emphasises chemistry, biological potential, and collection sites. A total of 115 compounds, including 30 new structures, were reported from 25 species across 17 genera. The reported findings demonstrated a broad spectrum of biological activities, including cytotoxic, antibacterial, antifungal, antiviral, anti-inflammatory, antimalarial, and antioxidant effects. However, the reported data and findings have stressed continued exploration of Red Sea organisms to support future drug discovery initiatives.

Flavonoids are bioactive plant secondary metabolites with various pharmacological activities. However, their application is limited by poor solubility, low bioavailability and instability under environmental conditions. Microparticle technology has emerged as a promising approach to overcome these limitations by enhancing flavonoid stability, solubility and controlled release. This narrative review summarises recent advances in the formulation and characterisation of herbal-based flavonoid microparticles, highlighting polymer selection, encapsulation techniques (e.g., solvent evaporation, ionotropic gelation, spray drying, freeze drying and spray chilling), physicochemical properties, and biological activities. The review emphasises that microparticle-based delivery systems may enhance flavonoid bioavailability and preserve bioactivity, offering a potential strategy for the development of novel therapeutic agents. These insights provide a comprehensive reference for future research on the pharmaceutical application of flavonoid microparticles.

Bhasmas are calcined metallic and mineral formulations used in traditional medicine and pharmaceutics that have attracted increasing scientific interest due to their nano-scale properties and potential pharmacological applications. However, their integration into mainstream pharmaceutical research remains limited by challenges related to standardization, safety validation, clinical evidence, and regulatory acceptance. This study presents a comprehensive bibliometric and thematic analysis of global Bhasmas research published between 1980 and 2024, with a pharmaceutical science–oriented perspective. Peer-reviewed publications indexed in PubMed and Scopus were retrieved using predefined Boolean search strategies, and a total of 407 eligible documents were analyzed using VOSviewer (version 1.6.20) to evaluate publication trends, collaboration networks, keyword co-occurrence, MeSH term clustering, citation patterns, and journal co-citation structures. The results indicate that India accounts for over 90% of global research output, with limited international collaboration involving the United States, the United Kingdom, and South Korea. Thematic analysis revealed a predominance of pre-clinical studies focused on metallic nanoparticles and animal models, with relatively few human studies. Significant gaps were identified in toxicological evaluation, pharmacokinetics, mechanistic understanding, and regulatory alignment. Overall, the findings highlight the need for interdisciplinary collaboration and adoption of modern pharmaceutical and analytical frameworks to enhance the translational relevance and global acceptance of Bhasmas-based formulations.

Given the limited number of new antibiotics in development and the decreasing effectiveness of those currently available, the miraculous impact of antibiotics in modern medicine appears to be nearing its end. Medicinal plants offer a promising alternative source for discovering new antimicrobial agents, given their vast diversity of biologically active constituents and the knowledge accumulated through centuries of use in traditional medicine.

In this review, we present a state-of-the-art overview of the antibacterial properties of five medicinal plants used to treat infections in ancient and medieval medicine and that continue to play a significant role in traditional therapeutic practices for addressing various human infections. The selected plants are Acorus calamus, Myrtus communis, and species of Boswellia, Urtica, and Plantago. The author conducted a comprehensive systematic search across PubMed, Scopus, and Google Scholar. The focus was on bioactive constituents, antibacterial susceptibility and spectrum, mechanisms of action, and suggested clinical applications in modern medicine for treating bacterial infections. This review synthesizes current knowledge, highlights the scarcity of pharmacological studies identifying medicinal compounds with specific antibacterial effects, and concludes that the development of new phytoantibacterial agents will depend on accelerating clinical research, particularly randomized controlled trials, to expedite their approval as new antibacterials.

The phosphatidylinositol 3-kinase (PI3K)–Protein kinase B (AKT)–mechanistic target of rapamycin (mTOR) signaling axis represents one of the most important oncogenic pathways in breast cancer and a major driver of therapeutic resistance, particularly in hormone receptor–positive, and triple-negative subtypes. Although PI3K and mTOR inhibitors such as alpelisib and everolimus have improved clinical outcomes in selected patients, their broader application is limited by toxicity and adaptive resistance mechanisms. Increasing attention has therefore been directed toward medicinal plant-derived phytochemicals as multi-target modulators of this pathway. This review summarizes current mechanistic evidence showing how selected phytochemicals—including flavonoids, polyphenols, and isothiocyanates—interfere with PI3K–AKT–mTOR signaling at multiple nodes, suppress compensatory survival pathways, and enhance the efficacy of endocrine therapy, chemotherapy, and targeted agents. Rather than acting as standalone therapies, these compounds are increasingly viewed as adjunctive modulators or lead scaffolds for anticancer drug development. This mini review uniquely integrates emerging evidence on phytochemical modulation of the PI3K–AKT–mTOR axis with mechanisms of therapeutic resistance and rational combination strategies with targeted therapies. It also highlights recent advances in nanoformulation technologies that may improve phytochemical bioavailability and facilitate clinical translation in precision breast cancer therapy.

Elimination of Enterococcus faecalis (E. faecalis) biofilms from the root canal system is a challenging issue in endodontics. The objective of this in vitro study was to evaluate the antibacterial efficacy of ultrasonic-activated herbal irrigants (black walnut [BW] and wormwood) and inorganic salts (alum and sodium chloride [NaCl]) against E. faecalis biofilms, compared with conventional irrigation. E. faecalis (MTCC 439) biofilms were grown on microtiter plates and treated with the test irrigants with and without ultrasonic activation for 5 and 10 minutes. Antibacterial efficacy was determined by Minimum Biofilm Eradication Concentration (MBEC) analysis using crystal violet staining and optical density measurement, and fluorescence microscopy using acridine orange and ethidium bromide staining to distinguish live and dead cells. All irrigants showed increased antibacterial activity when combined with ultrasonic activation (p < .05), with BW showing the highest biofilm reduction, followed by wormwood, alum, and NaCl. A 10-minute exposure time caused significantly greater biofilm disruption than 5 minutes. Ultrasonic activation dramatically enhances the antibacterial potential of herbal and inorganic irrigants, with BW, wormwood, and alum showing promise as adjuncts to conventional endodontic disinfection protocols.

The study aimed to develop and validate a precise, reliable, and sensitive LC–MS/MS method for the simultaneous estimation of Sofosbuvir (SOF), Voxilaprevir (VOX), and Velpatasvir (VEL) in rat plasma for pharmacokinetic applications. Plasma samples were protein-precipitated and analyzed using an Agilent Eclipse XDB C18 column with an acetonitrile–ammonium formate (20:80, pH 3.0) mobile phase, and detection was carried out on a SCIEX QTRAP 5500 in MRM mode. Optimized mass transitions were m/z 530.4537 → 294.1257 for SOF, 869.9463 → 259.6483 for VOX, 884.0635 → 761.3207 for VEL, and 883.0154 → 587.4326 for the internal standard, Elbasvir. The method demonstrated excellent linearity (R² > 0.999), acceptable precision (%CV ≤ 15%), and accuracy (85%–115%). Recovery of all analytes ranged from 95.39% to 98.74%, with negligible matrix effects. The LLOQ values were 20 ng/mL for SOF, 5 ng/mL for VOX, and 5 ng/mL for VEL. The developed method demonstrated acceptable accuracy, precision, and sensitivity for preclinical pharmacokinetic applications. The method was successfully applied to a rat pharmacokinetic study, indicating its suitability for exploratory bioanalytical investigations.

Indonesia produces abundant citronella essential oil, which contains diverse bioactive compounds with promising potential as natural feed additives, yet scientific evidence supporting its application in poultry nutrition remains limited. This study aimed to select the optimal and safest material composition for a self-nanoemulsifying drug delivery system (SNEDDS) of citronella essential oil using Design Expert Ver. 13.0.1.0, a D-optimal mixture, and to characterize the resulting formulation as a prospective poultry feed additive. Component selection, composition ratios, and oil phase proportions were determined using an exploratory approach: homogenization at 250 rpm for 15 minutes, followed by ultrasonication. The prepared SNEDDS formulations were evaluated through comprehensive physicochemical characterization, including organoleptic properties, percentage transmittance, emulsification time, dynamic light scattering (DLS) analysis, stability testing, and morphological observation. The composition ingredients of citronella SNEDDS consisted of virgin coconut oil (VCO) as the carrier oil, Tween 80 as the surfactant, and PEG 400 as the cosurfactant. The optimum formula consisted of 17.86% oil phase (citronella essential oil and VCO with a ratio of 50:50 [%]), 62.791% surfactant, and 19.169% cosurfactant. It demonstrated high transmittance (99.338% ± 0.366%), rapid emulsification (32.107 ± 0.454 s), small particle size (14.4 ± 0.141 nm), low polydispersity index (PDI) (0.088 ± 0.025), moderate zeta potential (–17.00 ± 2.980 mV), and appropriate viscosity (376.00 ± 3.637 mPa•s).

Antioxidant compounds present in plant-based foods and beverages play a crucial role in preventing oxidative stress by neutralizing reactive oxygen species. This study reports the formation and stabilization of the N,N,Nʹ,Nʹ-tetramethyl-p-phenylenediamine radical cation (TMPD^•+) in an aqueous medium and its application as a spectrophotometric probe for total antioxidant capacity (TAC) determination. Unlike the widely used ABTS^•+ and DPPH^•+ assays, which often require organic solvents, the proposed method operates entirely in water, representing a more environmentally friendly alternative.

The chemical parameters affecting TMPD^•+ stability were systematically optimized, including pH, buffer system, solvent effects, and oxidizing agents. The optimal conditions (λ_max = 563 nm) were established at pH 5.0 using Fe(III) 1.0 × 10^–4 M and TMPD 2.0 × 10^–4 M. Standard antioxidants were ranked according to their reducing capacity based on TMPD^•+ consumption, with tannic acid and ascorbic acid showing the highest responses. When applied to herbal teas, TAC values expressed as ascorbic acid equivalents showed a strong correlation with the ABTS^•+ assay (r = 0.900). These findings demonstrate that stabilized TMPD^•+ provides a simple, low-cost, sensitive, and greener approach for TAC determination in plant-based beverages.

Ixora coccinea, a medicinally important evergreen shrub, has been reported to exhibit numerous pharmacological activities, including neuroprotective effects. Hence, the current study aimed to investigate the molecular mechanisms and therapeutic potential of I. coccinea in managing Parkinson’s disease using integrated in silico and in vivo approaches. In a computational approach, bioactives of I. coccinea were screened and evaluated for their bioavailability. The potential targets regulated by bioactives were assessed via network, gene enrichment, and gene ontology analyses. Molecular docking was performed to predict the binding affinities, followed by molecular dynamics (MD) simulation. In experimental pharmacology, haloperidol was used to induce Parkinsonism in albino Wistar rats. Two doses of I. coccinea extract (200 and 400 mg/kg) were administered in conjunction with regular Syndopa. In the 21 days of treatment, rats were assessed for various neurobehavioral studies, biochemical, antioxidant and histological parameters. The network pharmacology revealed that stigmast-4-en-3-one showed the highest drug-likeness score of 0.91. Gene enrichment analysis predicted C-C motif chemokine 2, peroxisome proliferator-activated receptor alpha, and nuclear factor erythroid 2-related factor 2 (NFE2L2) as the top genes. Molecular docking revealed that NFE2L2, Catalase (CAT), and Peroxisome proliferator-activated receptor gamma (PPARG) showed the binding energy of −9.9, −9.5, and −9.3 kcal/mol, respectively. Molecular mechanics and MD simulation Poisson-Boltzmann study of surface area showed that the complex of CAT with 3-hydroxyflavone and PPARG with beta-sitosterol were most stable throughout the MD run. Further, I. coccinea ameliorated motor function, improved dopamine levels, reduced acetylcholinesterase (AChE) activity and oxidative stress, and displayed neuroprotection in the cerebral cortex region. The ethanolic extract of I. coccinea exhibited anti-Parkinson’s potential, possibly because PPARG and CAT are regulated by a variety of bioactives.

Tuberculosis (TB) remains a global health threat, with increasing resistance to first-line drugs like isoniazid necessitating the discovery of novel therapeutics. This study focuses on the in silico evaluation of 11 novel compounds of oxindole-triazole tethered derivatives (NV1-NV11) against the catalase-peroxidase enzyme of Mycobacterium tuberculosis (PDB ID: 1SJ2). Molecular docking was performed to evaluate binding scores, revealing that compound NV10 exhibited the strongest interaction with a high binding affinity of −10.262 (kcal/mol), showing a more favorable docking score than isoniazid (−6.331 kcal/mol). Molecular dynamics simulations (MDS) over 100 ns confirmed the structural stability and compactness of the NV10-1SJ2 complex, supported by stable RMSD, RMSF, and radius of gyration profiles. MM-GBSA binding free energy calculations further validated NV10’s binding energy, with a favorable ΔG_bind of −68.21 ± 3.10 kcal/mol, primarily driven by van der Waals and Coulombic interactions. These results suggest that NV10 is a computationally promising candidate for developing new anti-TB agents.

Despite the widespread medicinal use of Dillenia indica L. across Asia, a comprehensive comparative analysis of its various morphological parts, specifically the root, has remained largely unexplored. This research introduces a novel comparative methodology to define the distinct chemical and biological signatures of four plant parts (leaf, stem, unripe fruit, and root). Notably, extracts from the root and stem exhibited approximately 2× higher DPPH scavenging activity and nearly 25× greater α-glucosidase inhibition compared to those from the leaf and fruit. The root extract specifically demonstrated a 5× higher hydrogen peroxide (H₂O₂) scavenging capacity than all other extracts, which strongly correlates with its elevated glyceride, phenolic, and xanthone content. Conversely, the leaf extract displayed selective α-amylase inhibition, an activity potentially attributed to its specific alcohol and terpenoid contents. Furthermore, all extracts demonstrated selective cytotoxicity against colorectal cancer cells over normal fibroblasts, with leaf extract showing the greatest potency (IC₅₀ = 297.83 µg/mL at 72 h). These findings highlight the part-specific bioactivities of D. indica, supporting the rational selection of plant parts for developing safe, targeted functional foods and herbal products for preventive healthcare and chronic disease management.

Borrelidin is a pharmacologically active 18-membered macrolide with antibacterial, antifungal, anticancer, and antimalarial activities; however, its pharmaceutical development has been constrained by low fermentation yields. In this study, borrelidin production by an endophytic Streptomyces sp. isolated from the medicinal plant Zingiber zerumbet (L.) Smith was significantly enhanced through systematic medium optimization. A high-producing colony (BR7) was selected, and key nutritional variables were optimized using response surface methodology based on a Box–Behnken design (BBD). Statistical analysis revealed that borrelidin biosynthesis is governed primarily by strong quadratic and interaction effects among carbon and nitrogen sources rather than by individual nutrients. The optimized agro-based medium increased borrelidin concentration to approximately 83 mg/L, in close agreement with model predictions and representing a substantial improvement over the unoptimized process. The use of low-cost agricultural substrates supports the scalability and economic feasibility of this fermentation strategy. Overall, this work establishes a robust upstream bioprocess framework that supports the pharmaceutical development of borrelidin as a potential active pharmaceutical ingredient (API).

Helicobacter pylori (H. pylori) infection is commonly treated using antibiotics and proton pump inhibitors; however, drug resistance and adverse effects continue to reduce treatment effectiveness. In this context, the present study aimed to explore the Ayurvedic formulation Kaidaryadi Kashayam, traditionally indicated for gastrointestinal ailments, including H. pylori infection, as a potential source of phytochemicals with multitarget regulatory activity. Initially, the medicinal plants were authenticated through DNA barcoding. Subsequently, the phytochemicals identified from Gas Chromatography-Mass Spectrometry (GC-MS) analysis, along with those retrieved through literature mining, were subjected to virtual screening against 18 selected H. pylori targets using molecular docking. Out of 671 screened compounds, 25 phytochemicals exhibited interactions with multiple targets, and among these, five compounds, proanthocyanidin, phytomelin, drieline, riboflavin, and (2E,4E)-15-(1,3-benzodioxol-5-yl)-N-(2-methylpropyl) pentadeca-2,4-dienamide, displayed docking scores greater than the respective mean values. Notably, riboflavin demonstrated affinity toward ten targets, and prediction efficacy (PE) analysis revealed its highest total score (1,470) among all compounds. The binding stability of riboflavin with the target protein exhibiting the highest docking score (112) was further validated through a 100 ns molecular dynamics simulation. Collectively, these findings highlight the potential of Kaidaryadi Kashayam-derived phytochemicals in modulating multiple pathways of H. pylori pathogenesis, with riboflavin emerging as a particularly promising lead candidate for further experimental validation.

Endophytic fungi associated with the mangrove plant Ceriops tagal are a promising source of antibacterial compounds. This study aimed to isolate and identify endophytic fungi from C. tagal and evaluate the antibacterial activity of their secondary metabolites. Fungi were isolated from roots, branches and leaves using a direct plating method and cultured in potato dextrose broth. Crude extracts were prepared with ethyl acetate and tested against Escherichia coli, Staphylococcus aureus and Candida albicans using the disk diffusion method. Ten isolates were obtained, and all inhibited at least one test microorganism with inhibition zones ranging from 6.6 ± 0.55 mm to 19.1 ± 0.05 mm. Three isolates, ECT52, ECT57, and ECT63, showed strong activity with inhibition zones above 10 mm. Molecular identification based on ITS and LSU sequences identified them as Neopestalotiopsis saprophytica, Schizophyllum commune and Muyocopron laterale. Extracts of S. commune ECT57 showed the strongest activity against S. aureus with an MIC of 62.5 ± 0.96 µg/mL. LC-HRMS analysis of ECT57 extract revealed the presence of several dominant compounds, especially Cyclo(phenylalanyl-prolyl), 6-Quinolinecarboxylic acid and Caffeic acid. These findings indicate that C. tagal contains endophytic fungi that can synthesize antimicrobial compounds, which may have potential relevance for antibiotic development.

Mitragyna speciosa (Korth.), commonly known as kratom, contains a variety of secondary metabolites, with mitragynine, an indole alkaloid, being the most significant due to its analgesic properties. However, the limited commercial availability of mitragynine standards highlights the need for effective separation techniques. The complexity of kratom’s plant matrix makes selective isolation of mitragynine challenging. This study investigates the use of dummy template molecularly imprinted polymers (MIPs) as selective sorbents for isolating mitragynine. Colchicine was selected as a promising dummy template molecule for mitragynine. Prepolymerization studies, including association constant and Job plot analysis, identified methacrylic acid in methanol as the optimal component for synthesis. MIP synthesis was optimized using the bulk polymerization method with a template-monomer-crosslinker ratio of 1:3:20, at 80°C for 24 hours. Template removal was achieved through sonication, employing a 16-cycle methanol, and infrared spectroscopy confirmed successful template removal. MIP adsorption followed a pseudo-second-order kinetic model and a Langmuir isotherm. Kratom extraction was performed using the reflux method with methanol. Separation of mitragynine was conducted via column chromatography with MIP as the sorbent, resulting in enhanced mitragynine isolation. Quantitative analysis showed that the MIP method achieved an isolation efficiency of 36.7%, whereas preparative thin-layer chromatography (PTLC) yielded only 3.607%. Meanwhile, visual observations using Thin-Layer Chromatography (TLC) demonstrated that MIP produced a cleaner chromatogram compared to conventional PTLC. These findings suggest that MIPs offer a promising alternative to conventional separation techniques for isolating mitragynine from complex plant extracts.

The study assessed the effectiveness of a novel plant-based collagen booster (PBCB) formulation in enhancing collagen synthesis in human dermal fibroblasts, comparing it to commercial hydrolyzed collagen products. Dermal fibroblasts are vital for skin health as they produce collagen, maintaining skin elasticity and strength. The new formulation combines amino acids, vitamins, and active ingredients from specific plant extracts to boost collagen production. In vitro experiments showed that this formulation increased collagen synthesis by approximately 5.42-fold compared to vitamin C and outperformed bovine and marine collagen products by 3.24 and 1.66 times, respectively and 4.19 times better than the PBCB product, at a 2.5 mg/mL concentration. Overall, the preliminary findings suggest that this novel formulation may have potential for supporting skin health and anti-aging applications; however, comprehensive molecular analyses, in vivo studies, and clinical trials are required for validation.