The development of robust manufacturing processes for active pharmaceutical ingredients (APIs) requires considerably more than the successful execution of the target organic synthesis. In practice, reaction selectivity, competing transformation pathways, impurity formation, product isolation, and solid-state behavior are often tightly interconnected, ultimately determining whether pharmacopoeial-quality material can be consistently obtained. Despite their critical importance, these relationships are rarely discussed in the open literature because industrial process development is typically protected by proprietary know-how.
Droperidol represents an instructive model system for studying these challenges. Although its molecular structure appears synthetically straightforward, the preparation of pharmacopoeial-grade API is complicated by a series of coupled process phenomena, including competitive reagent consumption, structurally related impurities affecting downstream isolation, oxidative degradation during processing, and the influence of solvent selection on the final solid form.
In this work, these process vulnerabilities are examined through representative reaction pathways, while the corresponding pharmacopoeial impurities (Imp-C, Imp-D, and Imp-E) are prepared in analytically pure form to serve as mechanistic models and reference standards. Their structures were confirmed by NMR spectroscopy, mass spectrometry, HPLC, and LC-MS. Rather than presenting impurity synthesis as an isolated objective, the study demonstrates how the controlled preparation and characterization of representative impurities can support mechanistic understanding and impurity-control strategies during pharmaceutical process development.
Together, these findings illustrate how pharmacopoeial impurities can reveal process vulnerabilities that may remain hidden when API development is evaluated primarily through reaction conversion.
Parkinson’s disease (PD) is a progressive neurodegenerative disorder characterized by the loss of dopaminergic neurons, resulting in impaired motor function and reduced quality of life. Conventional oral therapy with Safinamide is associated with limitations, including first-pass metabolism and restricted brain delivery due to the blood–brain barrier (BBB). The present study aimed to develop and optimize a Safinamide-loaded nanostructured lipid carrier (NLC)-based intranasal film to enhance brain drug delivery. Safinamide-loaded NLCs were prepared using the melt emulsification–ultrasonication method, with glyceryl monostearate and oleic acid as the solid and liquid lipid matrices, respectively, and Poloxamer 188 and Tween 80 as surfactants. A three-factor, three-level Box–Behnken Design (BBD) was employed to optimize the formulation based on particle size, entrapment efficiency, and drug release. The optimized formulation (F8) exhibited a particle size of 178 ± 1.8 nm, entrapment efficiency of 88.0 ± 0.7%, drug release of 83.0 ± 0.8%, and drug content of 98.8 ± 0.4%. The optimized NLCs were incorporated into an HPMC-based nasal film, which demonstrated suitable thickness (0.17 ± 0.01 mm), high folding endurance (120 ± 2.4), near-neutral surface pH (6.1 ± 0.1), and rapid disintegration within 42 ± 1.1 s. In vivo pharmacokinetic studies in Wistar rats demonstrated enhanced brain exposure following intranasal administration. The optimized Safinamide-loaded NLC nasal film exhibited the highest brain Cmax (7.40 µg/mL), brain AUC₀–₂₄ (82.95 µg·h/mL), and Brain/Plasma Ratio (2.023) compared with oral Safinamide and the intranasal NLC dispersion. The optimized nasal film also showed a brain-targeting efficiency (BTE) of 256.32% and a direct transport percentage (DTP) of 61.0%, compared with 183.91% and 45.6%, respectively, for the intranasal NLC dispersion. These findings demonstrate enhanced brain targeting following intranasal administration, with the optimized NLC nasal film showing superior brain exposure and brain-targeting performance. Overall, the developed Safinamide-loaded NLC nasal film represents a promising non-invasive approach for enhancing brain drug delivery and may offer potential for improving the therapeutic management of Parkinson’s disease.
Rheumatoid arthritis (RA) is a systemic autoimmune inflammatory disorder of the joints. The therapeutic efficacy of conventional anti-arthritic drugs is significantly limited. Hence, we developed novel rutin-loaded pH-sensitive cationic nanoparticles (RT-pH-CNPs) based on a chitosan-L-methionine (CSM) conjugate. These nanoparticles enhance drug permeation through the stratum corneum, and their pH-sensitive characteristics aid targeted drug delivery to inflamed arthritic joints.
RT-pH-CNPs were prepared by the ionic gelation technique and optimized via Box-Behnken statistical design. Ex vivo permeation studies were performed using a modified Franz diffusion cell fitted with excised porcine ear skin. In vitro anti-arthritic activity was evaluated using egg albumin and bovine serum albumin (BSA) denaturation assays, along with the human red blood cell (HRBC) membrane stabilization assay. Further, the skin irritation test was performed using Wistar albino rats to evaluate the dermal safety of the developed formulations.
RT-pH-CNPs were successfully developed with an excellent particle size (212.4 nm), zeta potential (28.4 mV), and high drug entrapment efficiency (93.76 ± 0.696%), favoring their topical application in RA treatment. The optimized RT-pH-CNPs demonstrated pH-dependent drug release and approximately 2–fold greater permeation than free rutin suspension. Confocal studies revealed deeper penetration of rutin from RT-pH-CNPs. Furthermore, RT-pH-CNPs demonstrated enhanced anti-arthritic activity compared with free rutin and exhibited a non-irritant profile, suggesting their potential safety for topical application.
These findings suggest the potential of the developed RT-pH-CNPs for the management of RA due to their enhanced permeability, pH sensitivity, and better anti-arthritic activity.
Melasma is characterized by dark spots on the skin, particularly on the face, caused by hormonal changes, sun exposure, and genetic predisposition. Glutathione (GSH) and tranexamic acid (TXA) can help prevent this discoloration. This study aims to develop and validate an HPLC‑FLD method for simultaneous quantification of both analytes in complex matrices and to investigate the influence of the nano-emulsion type in their skin permeation.
A C18 column (15 cm × 4.6 mm, 5 μm) was used as the stationary phase, while a mixture of methanol and phosphate buffer (pH 6.7) served as the mobile phase in a gradient elution system. For fluorescence detection, pre‑column derivatization was performed using ortho‑phthalaldehyde and N‑acetyl‑L‑cysteine in borate buffer (pH 10.5). Reaction conditions and product stability were also evaluated. Two emulsions (o/w and w/o) were formulated and evaluated in terms of oil phase, droplet size and zeta potential. A Salt-Assisted Liquid-Liquid Extraction (SALLE) approach was optimized for nano-emulsions pretreatments. An ex vivo permeability study was performed using Franz diffusion cells and porcine skin.
The method showed excellent linearity (R² > 0.999), sensitivity (LODTXA = 3.9 ng/mL, LODGSH = 2.6 ng/mL), and reliability (%RSDTXA < 2%, %RSDGSH < 2.5%, %Recovery = 100 ± 4%). The SALLE procedure provided clean extracts and high %Recoveries. Tape stripping studies and permeation results showed that w/o emulsions significantly enhance TXA uptake and penetration, whereas GSH showed limited permeation regardless of formulation, consistent with both its physicochemical properties and its chemical instability under the experimental conditions.
These findings highlight the importance of selecting vehicle specific to each molecule to achieve optimal topical performance.
Diosmetin (Dio), a bioactive flavonoid with antioxidant and anticancer potential, has limited pharmaceutical applicability because of its poor aqueous solubility and low bioavailability. This study developed Dio-loaded folic acid (FA)-conjugated zein (Zn) nanoparticles surface-modified with amylopectin (AP) for potential colon-directed delivery. FA was conjugated to Zn via EDC/NHS-mediated coupling and characterized by ¹H NMR. Dio-loaded nanoparticles were prepared by antisolvent nanoprecipitation followed by AP coating at different concentrations and Zn/AP mass ratios. The optimized FA-Zn-Dio-AP formulation (Zn/AP mass ratio 2:1) exhibited a mean particle size of 234.5 ± 1.20 nm, polydispersity index of 0.16 ± 0.033, zeta potential of − 24.02 ± 1.35 mV, encapsulation efficiency of 94.51 ± 1.37%, and drug loading of 11.11 ± 0.32%. FTIR, DSC, and PXRD findings supported Dio incorporation and reduced drug crystallinity within the nanoparticle system. The AP-modified formulation showed improved stability under thermal, pH, ionic-strength, and refrigerated storage conditions. Sequential in vitro gastrointestinal release demonstrated limited Dio release in simulated gastric fluid (8.60%) and simulated intestinal fluid (28.13%), followed by greater release in simulated colonic fluid (66.96% at 24 h). The release profile showed the best fit to the Higuchi model (R² = 0.9882). FA-Zn-Dio-AP exhibited greater cytotoxicity against HCT-116 colorectal cancer cells than free Dio, with an IC₅₀ of 186.61 µg/mL, while retaining substantial DPPH radical-scavenging activity (83%). Overall, the findings support FA-Zn-Dio-AP nanoparticles as a promising platform for controlled, colon-directed delivery of Dio.
With the decline in the effectiveness of antibiotics due to microbial resistance, antimicrobial peptides such as Melittin have emerged as an emerging therapeutic option. This phase I study will evaluate the safety and maximum tolerated dose of topical Melittin spray in healthy volunteers.
This study evaluated the safety and tolerability of topical Melittin spray in healthy volunteers using a single ascending dose design. Thirty participants were randomly assigned to five groups of six in a block design into intervention and control groups. The evaluated concentrations were 8, 16, 32, 48, and 64 mg/L. Participants were monitored for 3 days after administration. The aim of the study was to determine the safety and maximum tolerated dose of Melittin spray.
During the study, no adverse events (mild, moderate, or severe) and no serious adverse events were observed among the volunteers (0/30; 95% CI: 0.0–11.6%). Also, the examination of local skin reactions showed that no skin irritation or sensitivity occurred in any of the participants (0/30; 95% CI: 0.0–11.6%). The spray formulation of melittin was well tolerated, and the highest tested concentration in this study was determined to be 64 mg/L, which is 4 times the minimum inhibitory concentration.
The antimicrobial peptide spray melittin has a favorable safety profile and is well tolerated by healthy volunteers when applied topically to intact skin. These promising results support melittin's clinical development as a novel infection therapy; however, further studies in patients with wounds, burns, or diabetic ulcers are required before broad clinical application.
This study has been registered at IRCT under identifier 20190924044863N1
High-throughput permeation assays are essential tools in drug discovery for identifying small molecules with favorable permeability, yet quantifying their transport kinetics across biomimetic membranes often relies on resource-intensive chromatographic methods. Here, we propose a simplified calibration-free endpoint absorbance workflow (POne) for rapid permeability screening in the 96-well PermeaPad® format.
Using a single plate-reader measurement and avoiding compound-specific calibration curves during primary screening, we evaluated permeation of 42 chemically diverse small molecules selected to span a broad physicochemical space relevant to passive membrane diffusion.
POne showed strong correlations with conventional calibration-based UV quantification (PCal, R2 = 0.85) and with LC–MS/MS-based (R2 = 0.82) methods. Using a pragmatic 10% POne threshold, POne and PCal yielded concordant permeability classifications in 89% of cases, demonstrating the usefulness of the method for rapid compound screening. Furthermore, we evaluated the transferability of the POne workflow to a mucus-coupled permeation model to further mimic the intestinal barrier. Although mucus-dependent alteration in permeation was observed for some compounds, these experiments showed that mucus-derived matrix interference restricts direct optical quantification, limiting the applicability of the method to a subset of compounds with suitable spectral properties.
While the need for a chromophore structure limits the POne method, it offers a cost-effective, robust alternative for early-stage screening workflows.
Albendazole (ABZ), a Biopharmaceutics Classification System (BCS) class II drug, exhibits dissolution-limited poor oral bioavailability (less than 5%), which reduces its therapeutic efficacy. Compared with conventional tablets, gummies improve pediatric acceptability by enhancing palatability and ease of swallowing.
The present study aimed to develop an emulsified ABZ-loaded oral gummies to enhance ABZ dissolution and pediatric patient acceptability.
Based on saturation solubility studies, oleic acid, Tween 80, and Transcutol® HP were selected as oil, surfactant, and co-surfactant, respectively to form ABZ-loaded preconcentrate. A ternary phase diagram was constructed to identify compositions exhibiting acceptable emulsification upon aqueous dilution, and the selected preconcentrate was evaluated for maximum ABZ-loading capacity. The ABZ-loaded selected preconcentrate exhibited efficient emulsification upon aqueous dilution, producing droplets with a diameter below approximately 600 nm. Upon dilution, the formed ABZ-loaded emulsion was subsequently incorporated into pectin-based gummies which were evaluated for texture, sensory acceptability, weight/drug-content uniformity, solid-state behavior (Fourier-transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), and powder X-ray diffraction (PXRD), in-vitro/pH-shift dissolution, and accelerated stability.
The emulsified ABZ-loaded optimized gummies showed acceptable textural characteristics, good weight and content uniformity, and satisfactory sensory acceptability with effective taste masking. Solid-state characterization by FTIR, DSC, and PXRD confirmed that ABZ was present in an amorphous or molecularly dispersed state within the optimized gummies. In vitro dissolution studies in 0.1 N HCl demonstrated significant improvement in drug release from the emulsified ABZ-loaded optimized gummies compared to pure ABZ (f2 < 50).
ABZ remained in an amorphous or molecularly dispersed state in the formulated gummies with no sign of recrystallization during accelerated stability studies (40 °C/75% RH).
Until now, no definitive treatment has been introduced for osteoarthritis. In this study, due to their unique characteristics, we intend to use the anti-inflammatory properties of postbiotics to control inflammation.
The K4IM cell line was cultured in a DMEM culture medium. After 24 h, 10 ng/ml TNF-α was added. Postbiotics were prepared with a standard protocol. Then, the optimal concentration of postbiotics was added to the cells. Finally, the cells were collected, their total RNA was extracted, and the cDNA was synthesized. The target genes’ mRNA expression level was checked by the qRT-PCR method. Pro and anti-inflammatory cytokines concentration was assessed using the ELISA technique.
In the cell line related to osteoarthritis, after crossing the cells with postbiotics extracted from yeasts, we had a significant decrease in inflammatory markers such as IL-1β, IL-6, IL-8, and CXCL-8 (p = 0.008, 0.013, 0.02, and 0.015, respectively), which resulted in gene expression levels being aligned. Meanwhile, anti-inflammatory markers such as IL-10, IL-4, and VEGF showed a significant increase in gene expression levels after the effect of postbiotics (p = 0.037, 0.045, and 0.041, respectively). IL-6 and IL-8 concentrations were decreased after the impact of postbiotics (p = 0.025 and 0.042, respectively), whereas IL-10 concentration increased (p = 0.038). Our investigation revealed a significant downregulation of miR-155 and miR-92a expression (p = 0.031 and 0.011, respectively), whereas miR-10a (p = 0.021) and miR-150 (p = 0.022) expression was upregulated after incubated with postbiotics.
Our results showed that the anti-inflammatory properties of postbiotics can control inflammation due to their unique characteristics.
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