The present investigation was carried out to develop a taste-masked, orally disintegrating tablet containing Dexketoprofen for evaluating the effect of the coating amount on the product’s quality attributes via Quality by Design (QbD) systematical roadmap. Dexketoprofen, S(+)-enantiomer of bitter taste ketoprofen, involves an arylalkil group which is the most frequently used analgesic in the management of acute and chronic pain. To overcome of bitter-taste of the active pharmacological ingredients should apply a taste-masking approach. For this purpose, the bitter taste dexketoprofen particles were coated with a pH-dependent methacrylates polymer in which one of the method of taste-masking approaches. The experimental design was enforced with a four-factor, three-level Box–Behnken method within the framework of response surface modeling (RSM). A ready to use matrix excipient, Eudragit RS 30D, dextrates, aroma, and tablet pressing force were chosen as independent factors, and were assessed on four dependent factors: dissolution rate, disintegration time, tablet hardness, and friability %. Our findings indicate that when tablet pressing force is applied at 250 PSI, the tablets disintegrate within 1 minute, and the friability value is under 1%. Disintegration time increases as the coating amount increases. However, the Pareto charts shows engrossingly that the dissolution rate is affected mainly by tablet pressing force in first, third, and fifth time points, and by matrix excipient and coating in the 10th, 15th, 20th, and 30th time points. It was concluded that the QbD study helped to understand how the coating amount and process variables impacted the dissolution rate, disintegration time, tablet hardness, and friability % of the Dexketoprofen orally disintegrating tablet (ODT) finished product. Full article

Polyethyleneimine (PEI) is a synthetic cationic polymer recognized as a non-viral gene carrier with high transfection efficiency. However, cytotoxicity issues limit its use. Pluronic block-copolymers conjugated with PEI have demonstrated promising results for multiparametric target gene/drug co-delivery in cancer with reduced side effects. The goal of this work was to synthesize and characterize a novel nanosystem Pluronic L121-PEI for gene/drug co-delivery. For this purpose, hydroxyl groups from Pluronic were activated using acryloyl chloride leading to the synthesis of a diacrylate intermediate, which was further conjugated with PEI. FTIR and ¹H-NMR spectroscopy were used for structural characterization. Particle size, polydispersity index (PDI) and zeta potential were assessed by dynamic and electrophoretic light scattering, respectively. A fluorescent pyrene probe was used to evaluate the critical micellar concentration (CMC). A hemolysis experiment was performed to estimate the in vitro biocompatibility of the nanosystem. FTIR analysis showed that Pluronic diacrylate was successfully synthetized as a new band around 1730 cm⁻¹ (C=O bond) appears. Its conjugation with PEI was confirmed by the presence of a band between 3380 and 3390 cm⁻¹ (N–H bond). ¹H-NMR results showed characteristic proton peaks from Pluronic (-CH3 at δ1.1 ppm) and from PEI (-CH2-CH2NH- between δ2.7–3.4 ppm) and the molar ratio Pluronic–PEI was 1:2. The nanoparticles’ hydrodynamic diameter was ca. 125 nm with a PDI below 0.250, and a charge nearby +30 mV. The CMC was around 50 μg/mL. The hemolysis ratio of a 5 mg/mL nanomicellar solution was less than 5%. A novel Pluronic L121-PEI was successfully synthesized, which was able to self-assemble in aqueous solution leading to the formation of biocompatible cationic polymeric micelles. Their small size is suitable for tumor-targeting and as they are positively charged they can be also valuable for gene delivery. Overall, this new nanosystem could be a promising multiparametric nanoapproach for gene/drug co-delivery in cancer therapy. Full article

In recent years, our knowledge of the biological effects of cyclodextrins has grown significantly. Cellular actions of cyclodextrins originate in their ability to form complexes with lipophilic biomolecules. Cyclodextrins can target different types of molecules according to their size, for instance, alpha-cyclodextrins form complexes with phospholipids, while beta-cyclodextrins can bind cholesterol or prostaglandin E2. Due to their interactions with the main membrane constituents, cyclodextrins can affect the barrier function of biological barriers or influence the function of membrane proteins. Nevertheless, cyclodextrins can enter the cells by endocytosis and affect the intracellular cholesterol storage. Based on these findings, 2-hydroxypropyl-beta cyclodextrin (HPBCD) received the orphan designation for the treatment of Niemann–Pick disease type C. The endocytosis of cyclodextrins works in different cell types and can be applied in the delivery of drugs into the cells. The tissue distribution and pharmacokinetics of cyclodextrins could be further characterized by imaging techniques. Radiolabeled HPBCD and randomly methylated beta-cyclodextrin (RAMEB) were recently used to study their in vivo behavior by positron emission tomography. Interestingly, RAMEB accumulation was detected in prostaglandin E2 (PGE2)-positive tumors. These findings can promote further research and the application of cyclodextrins in inflammation and tumor diagnosis or targeting. The presentation aims to give an overview of the main biological effects of cyclodextrins and the recent results of this research field. Full article

Multicomponent crystallisation is a widely studied technique in pharmaceutical chemistry to enhance physical properties of API’s such as solubility, stability and bioavailability without chemically modifying the drug moiety itself. Methods to produce multicomponent crystals are varied with solution crystallisation being the predominant method. Crystal morphologies also influence an API’s properties with needle shaped crystals dissolving slower and possessing poor flow properties compared to a more equant block shape. In this paper, we discuss the preparation of co-crystals and co-crystal salts of two poorly soluble drugs, pyrimethamine and diflunisal. In particular, we compare production of multicomponent crystals via cosublimation with the more common methods of mechanical grinding and solution crystallisation. Samples are sublimed on a laboratory scale from both ends of standard 15 × 160 mm test tubes sealed under vacuum with two heaters were used to equalize the sublimation rates of the components. We show that a range of multicomponent pharmaceutical crystals can be prepared that are not accessible via solution crystallisation, including polymorphs and ansolvates. In addition to binary systems, ternary crystals can also be obtained via this technique. Various diflunisal co-crystals crystallise as thin needles and we describe the use of tailor-made additives to obtain unprecedented morphology control of gas phase crystal growth. Finally, we discuss the formation of co-crystal salts in the absence of solvent. Salt formation was observed to occur during gas phase crystallisations in accordance with the pKa rule of 3 and modelling studies were carried out to understand the nature of proton transfer in these crystals in the absence of a solvent. Full article

(1) Background: Cyclodextrins are used in various areas due to their ability to form inclusion complexes and to modify some properties of the guest molecule. The characteristics that can be improved are related to the solubility of poorly water-soluble drugs, the stabilization of labile guests against the degradative effects of the environment (oxidation, light and heat), bioavailability, decreasing side effects, taste modification or odour elimination and controlling of drug release. Additionally, ibuprofen is a widely used nonsteroidal anti-inflammatory drug in treating pain and inflammation, but the long-term use of this drug has been associated with gastrointestinal side effects and nephrotoxicity. This led to the introduction of new compounds of ibuprofen with an improved profile. Aim. The research project combines two actual topics in the pharmaceutical area: developing new safer drugs and improving the pharmacokinetic and pharmacotoxicological profile through complexation with cyclodextrins. The main objective was to develop drug delivery systems based on cyclodextrins and new ibuprofen thiazolidin-4-ones as potential analgesic and anti-inflammatory drugs. (2) Methods: Thiazolidin-4-one derivatives of ibuprofen were included in β-cyclodextrin complexes by co-precipitation (1:1M) and lyophilization methods. The inclusion complexes were characterized using spectral methods such as infrared analysis (FTIR), NMR spectroscopy and phase solubility studies. The surface morphology was studied using scanning electron microscopy (SEM). (3) Results and Conclusions: Four inclusion complexes with β-cyclodextrin and new ibuprofen derivatives with a thiazolidin-4-one structure were obtained and characterized. These can confirm the theoretical premises for an improved pharmacological and safety toxicological profile and can continue with future studies (in vivo biological evaluation of pharmacokinetic, analgesic and anti-inflammatory profile). Full article

The majority of antibiotics display low oral bioavailability due to reduced solubility in water and/or inefficient permeability across biological membranes. Their efficiency is further limited by the frequent presence of polymorphic structures with different pharmaceutical activities. In this communication, we present our latest results on the development of organic salts and ionic liquids from fluoroquinolones (FQ-OSILs) as highly efficient ionic formulations of this family of antimicrobials. Ciprofloxacin and norfloxacin were used as anions and as cations, by combination with biocompatible organic counter-ions. In vitro bioavailability studies showed that all prepared FQ-OSILs presented higher solubility in water than the original drugs. All compounds were found to be isomorphic and with tailorable antimicrobial activity according to the cation–anion combination against Staphylococcus aureus, Bacillus subtilis and Klebsiella pneumoniae strains. Full article

Triple-negative breast cancer (TNBC) is a highly aggressive type of cancer with limited therapeutic options. However, this type of cancer cells has shown overexpression of folate receptors, which bind with folic acid (FA) with high affinity. This feature can be used for targeting of nanocarriers, such as liposomes. In order to further examine the potential of increased efficacy by targeting the folate receptor, we prepared folate conjugated liposomes (DSPC/Chol/PEG/DSPE-PEG-FA) and loaded them with doxorubicin (DOX), an anticancer drug. For this, we first synthesized and verified the conjugate between FA and PEG-lipid (FA-PEG-lipid). After that, liposomes were prepared with thin film hydration method followed by probe sonication. Three different types of targeted liposomes having different concentrations of FA-PEG-lipid in their membranes (0.1 mol%, 0.5 mol%, and 1 mol%) were evaluated for their cytotoxicity (DOX-loaded liposomes) on MDA-MB-231 (epithelial human breast cancer cells), 4T1 (murine mammary carcinoma cells), and MCF7 (Human breast cancer cells); the two first are TNBC cells and overexpress FA receptor, the third does not. Cytotoxicity results proved that increasing amounts of FA on the surface of liposomes results in enhanced antitumor activity of liposomal-DOX in the case of the cancer cells, which overexpress the FA receptor. Full article

Nano-hybrid formulations combine organic and inorganic materials in self-assembled platforms for drug delivery. Laponite is a synthetic clay, biocompatible, and a guest of compounds. Poloxamines are amphiphilic four-armed compounds and have pH-sensitive and thermosensitive properties. The association of Laponite and Poloxamine can be used to improve attachment to drugs and to increase the solubility of β-Lapachone (β-Lap). β-Lap has antiviral, antiparasitic, antitumor, and anti-inflammatory properties. However, the low water solubility of β-Lap limits its clinical and medical applications. All samples were prepared by mixing Tetronic 1304 and LAP in a range of 1–20% (w/w) and 0–3% (w/w), respectively. The β-Lap solubility was analyzed by UV-vis spectrophotometry, and physical behavior was evaluated across a range of temperatures. The analysis of data consisted of response surface methodology (RMS), and two kinds of machine learning (ML): multilayer perceptron (MLP) and support vector machine (SVM). The ML techniques, generated from a training process based on experimental data, obtained the best correlation coefficient adjustment for drug solubility and adequate physical classifications of the systems. The SVM method presented the best fit results of β-Lap solubilization. In silico tools promoted fine-tuning, and near-experimental data show β-Lap solubility and classification of physical behavior to be an excellent strategy for use in developing new nano-hybrid platforms. Full article

Wound healing treatments continue to be a medical challenge, since complications usually lead to chronicization and comorbidities. Natural inorganic ingredients such as clays have been demonstrated to exert useful activities in this regard. Hectorite is a smectite clay with desirable rheology due to its layered structure and remarkable swelling capacity. These properties make it an appropriate excipient for semisolid systems. Nonetheless, the biocompatibility of natural hectorite has been scarcely addressed; the majority of studies centered on synthetic or functionalized hectorites. The aim of this study was to prepare and characterize a hectorite/spring water hydrogel. The hectorite clay mineral was subjected to a solid-state characterization, while the hydrogel (HTgel@10) was evaluated in terms of rheology, pH and in vitro biocompatibility and wound healing. Results demonstrated that the hectorite possessed a remarkable purity (84% w/w of hectorite), very similar to that of similar pharmaceutical excipients. HTgel@10 showed a non-Newtonian, viscoplastic to pseudoplastic profile and a stable pH for 12 months. In vitro tests reported that the hectorite and the HTgel@10 were biocompatible (cellular viability ≥ 70%). Specifically, the hectorite used in this study was more biocompatible toward fibroblasts than Veegum^® HS. The in vitro wound healing test revealed that HTgel@10 was able to favor the wound closure. Therefore, hectorite/spring water hydrogels could be considered as potential wound healing formulations with remarkable stability and safety. Full article

Low dissolution rates of poorly soluble drugs are the factor afflicting their bioavailability. The aim of this study is to prepare a centrifugal spinning-based formulation of a poorly soluble drug, oxcarbazepine, for the improvement of dissolution rate and hence quick action. Sucrose-based microfibers of oxcarbazepine were prepared by a centrifugal melt spinning technique using a cotton candy machine. The prepared microfibers were characterized using Scanning electron microscopy (SEM), PXRD, Differential Scanning Calorimetry (DSC) and FTIR. The optimum formulation was molded into tablets and tested for in vitro drug release and in vivo pharmacokinetic studies using rabbits as test animals. The results indicated that the centrifugal spinning rapidly produced dissolving microfibers (diameter are <10 µm and dissolve in few seconds). In these fibers, ~20% oxcarbazepine was loaded, and both the yield and drug loading efficiency were improved by incorporating polyvinypyrolidine (PVP) in the formulations. The dissolution studies have revealed >90% of the drug was dissolved in just 2 min as compared with drug alone that shows only 15% dissolution at this time interval. XRD and DSC analyses have shown the amorphous state of the drug in the fibers while the FTIR analysis showed chemical stability of oxcarbazepine in the fibers. In vivo studies have revealed a 2 h reduction in tmax of drug in the rabbits treated with microfibers as compared with controlled group which was given pure oxcarbazepine. The study concludes the potential of the centrifugal spinning technique for the production of drug loaded fibers that can significantly enhance the dissolution rates of poorly soluble drugs and thus produce formulations for quick action of such drugs. Furthermore, the sucrose-based formulation can enhance the palatability with the intention of attracting pediatric patients. Full article

Ciprofloxacin (Cipro) is a broad-spectrum antibiotic used against both Gram (+) and Gram (−) bacteria. Its biological half-life is very short (4–5 h) and its conventional administration forms present a limited absorption efficiency. For this reason, the aim of this work was to study other administration strategies based on topical films. Sodium alginate (SA), a naturally occurring polymer, and a recombinant elastin-like polymer (rELP) produced by advanced genetic engineering techniques were evaluated as potential carrier systems. The films were obtained by the casting technique, adding the Cipro by direct dispersion in the polymer solution using 16.6% w/w rELP or 1.5% w/w SA. The in vitro release assays were performed at 37 °C in physiological solution and with orbital shaking at 90 rpm. Cipro concentration was determined by ultraviolet (UV) spectrophotometry at 276 nm. The release profiles were analyzed and adjusted using the Lumped model developed and validated by our research group. Pharmaceutical interest parameters were calculated and compared for both polymer-Cipro systems: the time required to reach 80% of the drug dissolved (t_80%), the dissolution efficiency (DE) and the mean dissolution time (MDT). The SA-Cipro platform released the 80% of the drug in 35 min, while this parameter was 209 min for the rELP-Cipro system. The MDT_80% was 8.9 and 53 min for the SA-Cipro and rELP-Cipro, respectively, while the DE, evaluated at 200 min, was 66.6 and 58.8 for each platform, respectively. These parameter values demonstrate that the rELP films were able to modulate the drug release rate and for the SA ones, release can be considered immediate. Therefore, both systems are promising strategies for the topical application of Cipro. Full article

The past few years have witnessed major developments in nanotechnology with great potential in powering new therapeutic tools for cancer management. Our goal in this study was to develop a biocompatible nanoshuttle for the efficient delivery of 5FU in colorectal cancer patients. Silk fibroin/Poly(ethylene glycol) nanoparticles (SF/PEG NPs) were obtained and further loaded with 5FU. These nanoshuttles were characterized in terms of morphological properties, size and size distribution, drug uptake and release potential as well as in vitro cytotoxicity potential screening. The SF/PEG + 5FU NPs cytotoxicity was determined on HT-29 cells after determination of the lethal dose 50 and targeted the evaluation of the cells viability, proliferation potential and migration and invasion potential. The inflammatory profile of RAW 264.7 macrophage cells was also determined by flow cytometry. The basic cytotoxicity screening revealed that the pristine SF/PEG NPs displayed good biocompatibility while the 5FU-loaded NPs induced cytotoxic effects on HT-29 cells. More, the 5FU-loaded SF/PEG NPs significantly reduced the migration and invasion processes as compared with the unloaded NPs. Lastly, we observed that the cytokine inflammatory profile was significantly altered after the treatment with the 5FU-loaded SF/PEG NPs as compared with the unloaded nanoshuttles. Full article

Electrospun biomaterials for in situ tissue regeneration are advantageous as drug delivery systems because they can be used to regulate the acute inflammatory response to establish a regenerative microenvironment. Neutrophils are the first inflammatory cells recruited to an electrospun biomaterial, and their release of neutrophil extracellular traps (NETs) and secretion of inflammatory and regenerative signals modulate the microenvironment around the biomaterial. In this work, chloroquine diphosphate, an anti-inflammatory drug shown to decrease NET release, was electrospun into polydioxanone (PDO) fibers for local delivery and immunomodulation of biomaterial-interacting neutrophils. Electrospinning was optimized so that the eluted concentration of chloroquine reached a previously reported, therapeutic concentration of 10 µM within one hour. Subsequently, acute neutrophil–template interactions were evaluated in vitro with freshly isolated human peripheral blood neutrophils for 3 and 6 h. NET release was quantified through detection of NET-derived myeloperoxidase with an ELISA, and the secretion of inflammatory and regenerative factors was quantified with an immunomagnetic multiplex assay. The results indicate that the chloroquine-eluting templates significantly reduced NET release within the first 3 h, but had no effect at 6 h, suggesting a therapeutic window for modulating acute NET release or temporal changes in cell viability. Additionally, the elution of chloroquine increased the secretion of regenerative factors HGF, VEGF-A, and IL-22 while suppressing inflammatory signals MMP-9 and IL-8 at 3 and 6 h, indicating a shift towards a pro-healing neutrophil phenotype. Together, these data suggest that chloroquine-eluting PDO biomaterials may modulate the acute neutrophil response from inflammatory to pro-healing, which may significantly enhance in situ tissue regeneration. Future work includes in vivo studies to evaluate long-term effects on tissue integration and regeneration in a physiological environment. Ultimately, these electrospun biomaterials may function as immunomodulatory drug delivery systems that regulate the neutrophil response and enhance the potential for in situ tissue regeneration. Full article

Osteoarthritis is frequently treated in veterinary settings with non-steroidal anti-inflammatory drugs (NSAID) such as carprofen (CP). Its action over the articular cartilage can be enhanced by increasing drug uptake into the cartilage, alongside its site of action, and anticipating its rapid distribution towards the bloodstream. A pharmacokinetic study to evaluate carprofen nanoparticles (NP) after intraarticular administration (IA) in rabbits was performed through a modeling allometric approach. The pharmacokinetic analysis of plasma profiles showed a rapid CP distribution outwards the synovial chamber but mainly remaining in plasma (Vc = 0.14 L/5 Kg), according to its high protein-binding. The absorption data modeling showed the occurrence of two different release–absorption rate processes after nanoparticle administration in the synovial space, i.e., a fast rate process causing a burst effect and involving the 59.5% of the total CP absorbed amount and a slow rate process, involving 40.5%. Interestingly, the CP burst effect inside the joint space enhances its diffusion towards cartilage resulting in CP accumulation in about three times higher concentrations than in plasma. In line with these results, the normalized-by-dose area under the concentration vs. time curve (AUC) values after IA were 80% lower than those observed after the intravenous. Moreover, the slower slope of the concentration–time terminal phase after IA administration vs. intravenous (IV) suggested a flip-flop phenomenon (0.35 h-1 vs. 0.19 h-1). Notably, CP clearances are predictive of the pharmacokinetic (PK) profile of CP in healthy humans (0.14 L/h/5 kg vs. 2.92 L/h/70 kg) although an over-estimation has been detected for cats or dogs (10 times and 4 times, respectively). This fact could probably be attributed to inter-species metabolic differences. In summary, despite the limited number of animals used, this study shows that the rabbit model could be suitable for a predictive evaluation of the release enhancement of CP-NP towards the biophase in arthritic diseases not due to sterical retention of the formulation. Full article

Intramuscular delivery is one of the main routes for DNA vaccines administration. However, it requires large amounts of the DNA to be administered and external stimulation to encourage the internalization of the DNA. In this work, we consider alternative routes for less invasive administration and develop drug delivery systems (DDS) for intranasal administration. Chitosan polyplexes using sodium tripolyphosphate (TPP) as a crosslinker were prepared using the ionic gelation method. Our method allowed preparation of nanoparticles with the size below 50 nm, which is at least two times lower than previously reported sizes. Moreover, despite the small sizes, we obtained DNA encapsulation efficiencies of about 70%. Parameters that may affect the encapsulation efficiency were investigated, including different TPP-chitosan ratios and concentrations of DNA. We found that encapsulation efficiency of DNA inside the particles decreases with the increasing TPP-chitosan ratio. Moreover, increasing the DNA concentration leads to a higher encapsulation efficiency. Small (<50 nm) chitosan nanoparticles hold enormous potential as DNA carriers due to their physiological barriers and subsequent internalization. Full article

Albendazole (ABZ) and benznidazole (BZL) are drugs with low solubility used in parasitic infections treatment. In this research, solid dispersion (SD) technology was used to enhance ABZ and BZL performance by increasing their dissolution rate and solubility. SDs were prepared by the fusion method, employing Poloxamer 407 (P407) as carrier to disperse 32 of BZL or 50% w/w of ABZ. Furthermore, physical mixtures (PM) of P407 and either ABZ or BZL were also prepared, and then SDs and PMs were characterized. Dissolution tests of SDs, PMs and commercial formulations (CF) of ABZ and BZL were carried out and dissolution profiles were analyzed with the lumped mathematical model, which allowed parameters of pharmaceutical relevance to be obtained. The results indicated that ABZ SD presented an initial dissolution rate (IDR) 21-fold and 11-fold faster than PM and CF, respectively, while the IDR of BZL SD was 2.5-fold and 4.5-fold faster than PM and CF, respectively. For BZL formulations, the time required to reach 80% dissolution of the drug (t_80%) was 4 (SD), 46 (PM), and 239 min (CF), while the dissolution efficiency (DE) at 30 min was 85 (DS), 71 (MF) and 65% (FC). For ABZ formulations, t_80% was 2 (SD), value not reached (PM) and 40 min (CF), while the DE at 30 min was 85 (SD), 36 (MF) and 65% (CF). The SDs developed notably increased the dissolution rate, in consonance with the values obtained from the pharmaceutical parameters, which could lead to faster absorption and, consequently, increase the bioavailability of these drugs. Full article

Poor solubility, erratic bioavailability and delivery challenges associated with gliclazide, which is commonly used in type 2 diabetes mellitus (T2DM) treatment, are overcome by exploring electrospun nanofibers technology. Employing emulsion electrospinning method with polyvinyl alcohol (PVA) alone and in combination with poly(d/l-lactide-co-glycolide) (PLGA), nanofibers were fabricated. Different concentrations of PLGA at 0.05%, 0.10% and 0.15% w/v were added to PVA to achieve a modified drug release profile to meet the typical physiological needs of T2DM, such as a faster drug release at meals followed by prolonged release to maintain constant plasma glucose level, which is highly desirable in T2DM management. Fabricated gliclazide-nanofibers were characterized by various studies, such as solubility, in-vitro drug release, drug release kinetic, scanning electron microscopy (SEM), differential scanning calorimetric (DSC), and Fourier transform infrared (FTIR) spectroscopy. GLZNF2, formulation of Drug:PVA:PLGA 0.1:10:0.05% w/v produced optimized gliclazide nanofibers. The optimized GLZNF2 nanofibers were incorporated into gelatin capsule for oral administration. SEM image of optimized formulation (GLZNF2) shows cylindrical shaped fiber, indicating gliclazide incorporated homogeneously in polymers with average fiber diameter 4.357 ± 0.83 µm. The solubility and dissolution rate of gliclazide nanofibers significantly improved compared to pure gliclazide. The gliclazide nanofibers produce a biphasic drug release profile, initial fast release, followed by prolonged release. Oral fabricated gliclazide fibers have tremendous potential as a drug carrier, and alternative technology for the improvement of solubility, dissolution rate, reduction in the dosing frequency and better blood glucose control could be explored in T2DM management. Full article

This short paper investigates the solubilizing ability of various cyclodextrins with efavirenz as well as the formation of solid inclusion complexes of efavirenz with β-CD and γ-CD. (1) Background: Efavirenz is a non-nucleoside reverse transcriptase inhibitor used as first-line treatment for adult and pediatric human immunodeficiency virus type 1 infection (HIV-1). Belonging to class II of Biopharmaceutical Classification System (BCS), efavirenz is poorly water-soluble. Inclusion into cyclodextrins is a possible strategy for increasing its solubility. (2) Methods: Solubility modulation was investigated by the phase solubility method; inclusion of efavirenz with β- and γ-cyclodextrins was attempted by co-dissolution with co-precipitation; the precipitates were studied by DSC, FT-IR, powder X-ray diffraction and optical microscopy. (3) Results: Solid state analysis of the precipitates shows evidence of separate recrystallization of β-cyclodextrin and efavirenz, whereas in the case of γ-cyclodextrin, a single new phase was observed. (4) Conclusion: Results show that the cavity of β-cyclodextrin is too narrow to accommodate efavirenz and only γ-cyclodextrin, the largest of native cyclodextrins, is able to form a true inclusion complex with this bulky guest. Full article

(1) Background: It is known that RNA in Na⁺ salt form has only immunomodulatory activity, and in acid form additionally acquires anti-inflammatory activity and in combination with D-mannitol acquires an even broader antiviral effect. The study aimed to study the ability of adenosinmonophosphat (AMP) and oligoribonucleotides (ORNs) in acid and salt form and in combination with D-mannitol to affect the conformation and fluorescence of interferon (INF) α-2b and to determine the energy parameters of these interactions. (2) Methods: spectroscopy (time pulsed and fluorescence), isothermal nanocalorimetry. (3) Results: AMP and ORNs in acid form and complex with D-mannitol bind more strongly to interferon α-2b than salt analogues. In the interaction of interferon α-2b and acid AMP and especially in complex with D-mannitol, the reaction occurs exothermically and change in conformational mobility INF by increasing the content of disordered regions. When INF α-2b interacts with salt AMP, the reaction occurs endothermically, and probably the salt form increases the conformational stiffness of INF α-2b. The greater efficiency of nonradiative energy transfer from INF α-2b to acid AMP has been shown, due to the closer distance between molecules. (4) Conclusions: AMP in acid form interacts more actively and increases the conformational mobility of INF, at a greater relative distance and with less Gibbs energy compared to the salt form, which probably causes the appearance of additional biological properties of acidic AMP. Full article

Arsonoliposomes (ARSL) constitute a particular class of liposomes that incorporate arsonolipids (ARS) into their membranes. ARSL realize selective toxicity to cancer cells; thus, they are an important tool in the treatment of cancer. Folic acid (FA) is widely used in targeted drug delivery due to its high affinity for the folate receptors that are overexpressed in cancer cell membranes. The aim of our studies was to develop novel triple-negative breast cancer (TNBC)-targeted ARSL by incorporating folic acid-conjugated polyethylene-glycol PEG-lipid (FA-PEG-lipid) into their membrane and loading them with anticancer drug doxorubicin (DOX). ARSL incorporating 0.1 mol% of FA-PEG-lipid were prepared and loaded with DOX, using the active loading protocol. They were characterized for their size distribution, zeta potential and drug entrapment efficiency (%). Their cytotoxic activity towards TNBC cell lines, particularly MDA-MB-231 (epithelial human breast cancer cells) and MCF7 (human breast cancer cells), was evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide MTT-assay. The first results demonstrated enhanced toxicity of this novel type of ARSL towards cancer cells, which is particularly interesting and deserves further exploitation. Full article

Montelukast sodium (MLK) is a worldwide antiasthmatic drug. Commercial formulations still have some issues with solubility and instability to light and humidity. To overcome them, the present work reports inclusion compounds of MLK and γ-cyclodextrin (γ-CD). As a molecular capsule, CDs have the ability to protect the inclusion guest from degradation, enhance its solubility and alter the pharmacokinetic parameters. MLKγ-CD inclusion compounds were prepared by mechanochemistry. Without using any solvent, γ-CD was pre-milled and then co-milled with an equimolar quantity of MLK in a ball mill at 600 cycles·min⁻¹. After 120 min of milling, the formation of MLK·γ-CD inclusion compounds was confirmed by powder X-ray diffraction and scanning electron microscopy. Additional studies, performed under pharmacopeia guidelines, showed that the prepared MLKγ-CD inclusion compounds can indeed increase the dissolution of MLK when in ultrapure water or simulated intestinal fluid (without pancreatin). This way, the MLKγ-CD inclusion compounds that are presented in this work are a promising solution for improving the therapeutic effectiveness of MLK. Full article

Recent scientific publications have already demonstrated that co-grinding appears to be an efficient, solvent-free technique for preparing cyclodextrin (CD) complexes and improving physicochemical properties (solubility, stability, etc.) of active ingredients. The improvement of solubility and dissolution rate could enhance the biopharmaceutical properties of the active ingredients in pharmaceutical products. In this study terbinafine hydrochloride (TER), an antifungal BCS II drug was chosen as a model drug. The aim of this study was the follow-through of inclusion complex preparation by co-grinding, using several analytical methods. TER and two amorphous cyclodextrin derivatives (hydroxypropyl-β-cyclodextrin (HPBCD); heptakis-(2,6-di-O-methyl)-β-cyclodextrin (DIMEB)) were used for the preparation of products. Products were analyzed by differential scanning calorimetry (DSC), X-ray powder diffractometry (XRPD), hot-humidity stage X-ray powder diffractometry (HOT-XRPD), Raman spectroscopy, and Fourier-transform infrared spectroscopy (FT-IR). Dissolution studies of TER and products were also performed out. DSC and XRPD studies suggested that the crystallinity of products gradually decreased by the increasing grinding time, and after 75 min of co-grinding the products were completely amorphous. HOT-XRPD studies revealed that the amorphous product containing HPBCD did not change with increasing temperature. However, in the same process, the DIMEB-containing product recrystallized in a new crystalline phase. Raman and FT-IR spectroscopy confirmed the molecular interactions between the components. Dissolution studies showed that the dissolution rate of complexes improved, and the solubility of TER increased in both simulated gastric and intestinal fluid, depending on the pH of the dissolution medium. Full article

Joint pain with high prevalence and yet without any specific treatment option is posing a challenge to healthcare professionals day by day. Amongst several treatment options currently utilized for arthritic joint pain are merely giving symptomatic relief rather than curative treatment. Nonsteroidal anti-inflammatory drugs (NSAIDs) are the most widely accessed treatment option amongst all of the options. However, their adverse effects profile is a major hurdle for their use, especially in elderly patients. The present study was focused on developing a transdermal patch of a novel NSAID Loxoprofen sodium with enhanced penetration and improved patient compliance. Pluronic lecithin organogel (PLO) was selected as transdermal drug delivery platform to enhance its penetration through skin. Moreover, the transdermal route bypasses first-pass metabolism, gastrointestinal (GI) side effects, and the necessity to administer drug orally. All of these credentials ultimately improved patient compliance. Several experimental batches (PL1 to PL8) were formulated to prepare the PLO of loxoprofen sodium. All the batches were evaluated for physical appearance, pH, viscosity, spreadability, drug content, and in vitro drug diffusion profiles. An optimized batch was selected on the basis of the obtained results. It showed sustained drug release up to 12 h. The study evidenced that similar transdermal formulations of other NSAIDs can significantly enhance current treatment scenario for joint pain. Moreover, conversion of such formulations in transdermal patches or other forms ensure sustained and reproducible transdermal flux, which can be further fabricated as bioequivalent to the oral formulations. Further studies can be designed to evaluate the clinical applicability of the formulation. Full article

Many neuroactive drugs are characterized by poor solubility, hampering their therapeutic potential and clinical research studies. For instance, the lipophilic molecules dimethylfumarate, retinyl palmitate, progesterone, and URB597 can be employed in the treatment of relapsing remitting multiple, early brain injury, learning deficits, and/or traumatic brain injuries. In this study, the possibility to encapsulate these drugs in lipid nanoparticles is investigated. Solid lipid nanoparticles and nanostructured lipid carriers have been produced by melt and ultrasonication of stearic triglyceride or a mixture of stearic triglyceride and caprylic/capric triglycerides. Mean diameters and morphology of lipid particles were studied by photon correlation spectroscopy, cryo-transmission electron microscopy, and x-ray diffraction, while encapsulation efficiency and in vitro drug release have been determined by HPLC. A behavioral study was conducted in rats to study the capability of lipid nanoparticles containing URB597 to alter behaviors relevant to psychiatric disorders after intranasal administration. In this regard, the nanoparticle surface has been modified by polysorbate 80 in order to obtain “stealth” nanoparticles. The nanoencapsulation strategy allowed increased drug solubility with respect to unphysiological solvent or solvent mixtures usually employed for animal and clinical studies. In particular, retinyl palmitate solubility in nanostructured lipid carriers has been increased up to eight-fold. Moreover, rat behavioral effects observed by nanoencapsulated URB597 administered intranasally suggest the therapeutic potential of this non-invasive route to treat social dysfunctions, such as autism. Full article

Iloperidone (ILO) is a second-generation antipsychotic drug and a first-line treatment approved by USFDA in May 2009. Iloperidone belongs to Biopharmaceutical Classification Systems (BCS) class II; thus, it is poorly water-soluble, highly permeable, and has pH-dependent solubility. Cyclodextrins and their derivatives have a wide range of applications in different formulations due to their complexation ability, which improves the solubility, stability, safety, and bioavailability of a drug. We have tried the complexation of iloperidone with sulfobutyl ether-β-cyclodextrin (SEβCD) to improve its solubility and dissolution. Complexation was done by the kneading method. The characterization of the SEβCD complexes with Iloperidone was done by FTIR, differential scanning calorimetry (DSC), saturation solubility, etc. A multimedia dissolution of the complex was carried out and compared with the plain drug. A significant improvement in drug release was found from SEβCD complexes in all media when compared with the drug alone. Full article

Interest has developed in natural molecules due to their clinically proven effects on skin diseases. Flavanones display several biological activities, and recently have been the focus of studies due to their anti-inflammatory effect. To improve their pharmacological profile, four flavanones (A, B, C, and D) were synthesized by structural modification of one natural flavanone 1 (semi-systematic name: (2S)-5,7-dihydroxy-6-prenylflavanone) extracted from Eysenhardtia platycarpa. The hydroalcoholic flavanone solutions (FS) were assayed to investigate their anti-inflammatory effect on two in vivo cutaneous inflammation models. Materials and methods: the topical anti-inflammatory effects of FS were evaluated against models of 12-O-tetradecanoylphorbol acetate (TPA)-induced mouse ear edema and arachidonic acid (AA) in rat ear edema. Results: The vinylogous cyclized derivative (flavanone D) caused edema inhibition in the TPA-induced models with an inhibition of 96.27 ± 1.93%; equally effective and potent in inhibiting the mouse ear edema as indomethacin had been. In addition, the AA-induced increase in ear thickness was reduced the most by the topical application of modulated ether (flavanone B). Conclusions: The in vivo and histology results suggest that flavanones B and D are effective as topical anti-inflammatory agents in inflammatory processes. Thus, this new compound represents a promising agent for the management of skin diseases with an inflammatory component. Full article

Among the most promising technologies for sustained drug delivery systems are core-shell nanofibres prepared by electrospinning. However, the most common method for the production of those, coaxial electrospinning, suffers from extremely low flow rates limiting the practical applications of such fibres. Emulsion electrospinning, on the other hand, enables the use of the high-throughput needle-less electrospinning devices for the production of the core-shell nanofibres with active pharmaceutical ingredients embedded and protected in their core. The development of such drug delivery systems for the applications in bone regeneration is further challenged by the need for the inorganic additives meant to stimulate the regeneration of the bone. The main objective of this work is to develop a high-throughput electrospinning method for the production of hybrid (organic-inorganic) and bioactive scaffolds needed for bone regeneration. We demonstrate the importance of the formulation, e.g., presence of surfactants, on the stability of the emulsion, and thus on electrospinning. Our work is an important step forward towards the high-throughput production of complex multi-material scaffolds for sustained drug delivery. Full article

Topical application of flavonoids has recently received increased attention, however their use is limited due to a low aqueous solubility and related low in vivo absorption. Monolein emulsified in water leads to aqueous nanostructured dispersions of complex lyotropic liquid crystalline phases able to carry lipophilic molecules. These monoolein aqueous dispersions (MADs) were investigated on two model flavonoids, namely quercetin and rutin. MADs were produced by emulsifying monoolein in water in the presence of sodium cholate. MAD size, morphology and drug content were characterized using PCS, SdFFF, cryo-TEM and UV spectroscopy. In vitro studies on drug release and antioxidant activity were also conducted. MAD size was found around 300 nm. Cryo-TEM showed that sodium cholate content influences the morphological aspect of MAD. Concerning drug content, MAD increased at least 80-fold quercetin solubility, while the same was not found for rutin. Experiments on antioxidant activity demonstrated the two-fold power of quercetin as compared to rutin while in vitro Franz cell experiments showed that MAD are suitable for cutaneous application. In conclusion MAD can be potentially proposed for the delivery of the antioxidant molecule quercetin, whilst many studies have to be performed for finding a way to deliver rutin. Full article

Amphotericin B (AmB) is a high-molecular weight poorly soluble drug with a high efficacy in the treatment of infectious caused by Leishmania spp. parasites, which possesses a very low topical bioavailability. Transferosomes (TFs) are ultradeformable vesicles that consist of drugs lipids, an edge activator, and a low amount of ethanol (<10%). They have been engineered and optimized to enhance the permeation of AmB across the skin and, thus, its antiparasitic activity. Drug loading of the formulation resulted in 0.086%, while a good physicochemical stability for 6 months under desiccated conditions was observed. AmB-TFs illustrated a flux of 4.91 ± 0.41 µg/cm²/h across mouse skin. In vivo studies demonstrated a good permeation of the drug after topical application on healthy mouse skin, which was increased using microneedles at early exposure times, while in vivo efficacy studies demonstrated that the parasite load was decreased in a 98.24 ± 1.54%. Full article

Most new low molecular weight chemical entities in pharmaceutical developments suffer from a low aqueous solubility, making oral delivery challenging. Despite the numerous formulation efforts that can be investigated, research especially on amorphous drugs and formulations appears to be a useful approach. Whilst few drugs can be converted to an amorphous form on their own, due to their physical stability, the use of amorphous solid dispersions, i.e., the dissolution of drug molecules into (amorphous) polymers is increasingly used. However, certain shortcomings of these polymers based amorphous solid dispersions, such as a low drug load and a usually high hygroscopicity, still necessitate the investigation of alternative approaches. One such approach is the use of co-amorphous systems, i.e., the combination of initially crystalline low molecular weight drugs and excipients. Usually, here, a 1:1 molar ratio is used, but this may not be the optimal mixing ratio. In this presentation, work on investigating the optimal ratio between drug and co-former will be presented and critically discussed. Full article

Cancer is one of the leading causes of death in the world and protein therapeutics play an important role in combating this disease. Novel nanocarriers are needed for optimal delivery, enhanced therapeutic effect, and protection of proteins. Poly Lactic-co-Glycolic Acid (PLGA) nanoparticles are commonly used, since they are non-toxic, biodegradable, and allow for the sustained release of the active pharmaceutical ingredient (API). Accurate quantification of the therapeutic inside these nanocarriers is essential for further development and precise in vivo experiments, especially for determining the correct therapeutic dose. Bicinchoninic acid (BCA) assay is one of the most popular methods of protein quantification, known for its low sensitivity to common surfactants. However, large discrepancies between published results are often observed, with determined protein encapsulation efficiencies (EE) varying from 20 to 80%. We investigate the interference of excipients or the combination of excipients, on accurate EE determination of PLGA nanoparticles using the micro BCA assay. The EE was determined using multiple methods: by measuring the un-encapsulated protein (indirect approach) and directly by extracting the protein using sodium hydroxide and dimethyl sulfoxide. We show differences between the methods, highlight the most common pitfalls, and show the importance of using correct standards in assessing EE. Full article

Cancer is the second leading cause of death in the world and is often untreatable. Protein-based therapeutics, such as immunotherapeutics, show promising results in the fight against cancer, resulting in their market share increasing every year. Unfortunately, most protein-based therapeutics suffer from fast degradation in the blood, making effective treatment expensive, causing more off-target effects (due to the high doses necessary), and often require repeated injections to stay within the correct therapeutic range. Encapsulation of these proteins inside nanocarriers is prompted to overcome these problems by enhancing targeted drug delivery and, thus, leading to a less frequent administration and lower required dose. However, most current protein encapsulation methods show very low loading capacities (LC). This leads to even more expensive treatments and might pose a further risk for the patient caused by systemic toxicity against high concentrations of the carrier material. We investigated and optimized protein nanoprecipitation as a method to obtain a high protein LC and encapsulation efficiency (EE) inside poly(lactic-co-glycolic acid; PLGA) nanoparticles via a simple two-step process. In this work, we used model proteins to investigate the influence of various parameters such as precipitation solvent, addition speed, and protein concentration on protein activity. Our work is a critical step towards the high-loading encapsulation of immunotherapeutics. Full article

Cannabidiol (CBD) has emerged as a potential therapeutic agent. However, its low aqueous solubility hinders the development of effective parenteral formulations. The use of polymeric microparticles as CBD carriers could resolve this challenge and allows to obtain an extended CBD release after a single administration. Poly(lactic-co-glycolic acid) (PLGA), FDA approved for various medical applications, is one the most used. Ionizing radiation is as an effective sterilizing method for PLGA microparticles, which is essential for their parenteral administration. The aim of this work was to evaluate the effect of gamma sterilization on the characteristics of CBD-loaded microparticles. No differences in particle morphology and particle size were detected between sterile and non-sterile formulations. All microparticles exhibited a spherical shape, a smooth surface, and an average particle size around 25 µm. DSC analysis showed the absence of the CBD melting peak in sterile and non-sterile CBD-microparticles. However, a reduction on PLGA glass transition was appreciated in sterile formulations. A significant lower CBD content was also detected in sterile microparticles, indicating a CBD degradation during sterilization. Finally, a faster CBD release was appreciated in sterile microparticles compared with their counterparts, due to the faster PLGA degradation in sterilized microparticles. Due to the CBD degradation during sterilization process and the acceleration of the release of this drug from PLGA microparticles, gamma irradiation is not an adequate method to sterilize CBD-PLGA-microparticles. Full article

Self-emulsifying systems (SEEDS) are becoming increasingly popular for the preparation of oral dosage forms of low-water-soluble drugs. SEEDS are divided into subgroups according to their components; type IV formulations that are prepared using only surfactants and/or co-surfactants. The aim of the study was to increase the solubility of tadalafil, a class II drug according to the BCS, by preparing a type IV formulation. In the study, Labrasol, Kolliphor PS 20, Kolliphor PS 60, Kolliphor PS 80, Kolliphor CS 12, Kolliphor CS 20, Kolliphor HS 15, Kolliphor EL, Kolliphor ELP, Kolliphor PEG400, Gelucire 44/14 and Gelucire 48/16 were used as surfactants; Transcutol was used as a co-surfactant. The Kolliphor PS 80, Kolliphor EL, and Kolliphor HS 15 formulations were prepared in a ratio of 2:1 with Transcutol to form droplet sizes less than 50 nm and PDI values below 0.2. The formulations sensitivity to heat change and pH change was analysed by performing stability tests. Stable formulations of tadalafil formulations were obtained using Transcutol and Kolliphor PS 80 or Kolliphor EL. Solidifications of the optimum type IV formulations were made using fluorite, Neusilin US2, Neusilin FL2, Syloid 3050, and Syloid 3150. Dissolution studies of the prepared solid type IV formulations were first performed in 0.1 N HCl, and the dissolution profiles were examined by analysing the optimum solid type IV (s-type IV) formulations in pH 4.5 and pH 6.8. It was determined that the s-type IV prepared with Neusilin US2 and Neusilin UFL2 (2:1) had provided a dissolution of over 80% at the end of the first minute. The results indicated the potential bioavailability improvement of s-type IV for the BCS class II drug, Tadalafil, due to great enhancement in its dissolution rate. Full article

Electrospinning is one of the most promising techniques for producing nanofibers for pharmaceutical and medical applications. Electrospun fibers are especially promising as sustained release drug delivery systems of biotherapeutics. However, a typical electrospinning system made of a single needle configuration has a low fiber production efficiency and throughput. Good candidates to overcome the low productivity issue are the needleless spinnerets. Many of them have already been experimentally and theoretically tested, but their comprehensive comparison is still needed. This study summarizes the most efficient needle-free electrospinning configurations based on their extensive electric field profile description simulated using the finite element method (FEM). Understanding these properties is crucial when choosing the electrospinning system that allows high-throughput production of the monodispersed nanofibers. The ability to control the process will enable its applications in drug delivery systems. Full article

Rosuvastatin calcium (Rsv) is an effective statin, with a potent antihyperlipidemic effect. However, it suffers poor bioavailability owing to its poor solubility. Thus; encapsulating Rsv into a nanovesicular structure could overcome this problem. The aim of this work is to investigate the potential of solid lipid nanoparticles (SLNs) and nanostructured lipid carriers (NLCs) in enhancing the solubility of Rsv, using the quality by design (QbD) concept. A complete risk assessment study has been conducted, where the critical process parameters (CPPs), material attributes (MAs), and critical quality attributes have been identified using Ishikawa diagrams. Selected CPPs/MAs were screened and further upgraded to a 2⁴ full-factorial design to develop a design space with the optimized formula. The screened CPPs/MAs were tested on the particle size, the polydispersity index (PDI), the zeta potential (ζ-pot), and the entrapment efficiency (EE%). A comprehensive approach for Rsv nanovesicular carriers has been conducted, where the NLCs showed better results than the SLNs. The optimized formula was prepared with 3% total lipid content, 0.154% surfactant, and 9.4 mg drug. The optimized formula had a particle size of 310.5 nm, with 0.243 PDI, a ζ-pot of −24.7 mV, and an EE% of 93.87%, and showed a sustained release of the drug for up to 72 h. It successfully lowered total cholesterol, low density lipoprotein, and triglycerides, and elevated the levels of high density lipoprotein in rats, with better results as compared to the standard drug. Thus, a complete QbD study was conducted to explore experimental regions for many successful nanovesicular carriers for the enhancement of the solubility of poorly soluble drugs. Full article

Pre-stability studies carried out throughout the development of a diclofenac emulgel formulation have shown a clear decrease in the drug release rate. In order to address the root-cause associated with this phenomena, product historical data were retrieved and analyzed following a retrospective Quality by Design (rQbD) approach. The quality target product profile (QTPP) was established, and risk assessment tools were used to identify the most relevant parameters affecting formulation performance. These consisted in (i) mixing time, (ii) sodium hydroxide content and (iii) carbopol grade. Following a 2³ full factorial design, the pH, viscosity, in vitro release rate and cumulative amount of drug released at the end of the release experiment were selected as responses to statistically model the available data. It was observed that higher sodium hydroxide concentrations induce a decrease in viscosity, consequently resulting in a superior pharmaceutical performance. Moreover, as a secondary effect, a lower carbopol viscosity yields lower release outputs. The estimated models were used to define a feasible working region, which was further confirmed at an industrial scale. This work highlights the use of rQbD principles to achieve a greater product understanding. By doing so, specific strategies can be applied to product manufacture in order to consistently meet QTPP requirements. Full article

Thermosensitive in situ gels are promising formulations for the management of Alzheimer’s disease (AD), since they increase the residence time of lipid-based nanosystems in the nasal cavity, improving drug therapeutic efficacy. The purpose of this study is to prepare thermosensitive in situ gels with anticholinesterase inhibitor (RVG)-loaded nanostructured lipid carriers (NLC) and nanoemulsions to improve the residence time of the formulations in the nasal cavity. Different concentrations of thermosensitive polymers were added to the RVG-loaded NLC and to the RVG-loaded nanoemulsion to optimize the gelation temperature of the in situ gels; concentrations of 17% (%, w/w) of Kolliphor^® P407 and 0.3% (%, w/w) of Methocel^TM K4M were selected. The in situ gels of the RVG-loaded NLC and RVG-loaded nanoemulsion had a particle size, PDI, ZP, and pH of, respectively: 141.70 ± 0.40 nm and 146.10 ± 1.73 nm; 0.45 ± 0.00 and 0.43 ± 0.02; −4.06 ± 1.03 mV and −4.09 ± 0.71 mV, 6.60 ± 0.01 and 7.00 ± 0.02. In addition, these in situ gels showed a non-Newtonian plastic behavior, and the texture parameters presented desirable values for nasal administration. From these results, we concluded that the developed in situ gels can be used to improve the treatment of AD through the nose-to-brain route. Full article

Optimization of transdermal formulations requires solving simultaneous challenges as the selection of release polymers. The interactions between the formulation components must be taken as a way to modulate its performance. Selection of acrylic polymers with different functionalizations for the transdermal formulation of a tertiary amine drug (ropinirole HCl) have been investigated. Aim of this work is to characterize the influence over drug release of certain experimental interactions. Solubility-crystalization and pharmacopoeial release tests have been used to evaluate the influence of drug loading and the pH of the release media. Area under the curve of dissolved amounts and percentage of release have been used as discriminant variables in mutual influence with the physical state of the drug. Elucidation of release mechanisms has been performed with data fitting of relevant modelystic equations. Fickian release and erosion contribution have been related with drug loading and the risk of burst effects. In conclusion, a rationale to select the best suitable polymer for ropinirole HCl has been demonstrated in terms of efficiency and extent of release. Full article

High-throughput emulsion electrospinning is a technology that can enable practical nanofiber application for drug delivery. Core-shell structure of the electrospun fibers allows the encapsulation of the active pharmaceutical ingredients (APIs), protects their activity, and controls their release rate. However, electrospinning using high flow rates usually requires high electric fields that may negatively affect the activity of the biomolecules. Moreover, charged APIs tend to migrate to the surface of the fibers during the electrospinning process leading to the high burst release. That is disadvantageous when long-term sustained release is needed. We have investigated the influence of the electrospinning parameters such as distances between the electrode and collector and the applied voltages to both activity of the encapsulated proteins and their burst release. We have also tested the influence of number of the stabilizers, e.g., trehalose, pluronic, and polyvinylpyrrolidone, on their ability to preserve the protein activity, and the influence of the different molecular weights of polyvinyl alcohol on the ability to sustain the release. Our results demonstrate the importance of the water phase composition to both activity and release and are critical for further understanding of the processes taking place during the emulsion electrospinning. Full article

The intranasal administration of nanostructured lipid carriers (NLCs) has been suggested as a promising strategy to improve the fast treatment of epilepsy. This route allows for drug passage directly from the nose to the brain, avoiding the need of bypassing the blood–brain barrier. In addition, the quality-by-design (QbD) approach is a useful tool for the optimization of manufacturing variables, resulting in effective and safe pharmaceutical formulations. The aim of this work was to use the QbD approach to optimize a NLCs formulation for the nose-to-brain delivery of diazepam. The studies began with the screening of excipients and the assessment of the lipid-drug compatibility. The central composite design was used to evaluate the effects of critical material attributes (CMAs) (ratio of solid and liquid lipids and the amount of drug and emulsifiers) on the CQAs of the diazepam-loaded NLCs formulation (particle size, polydispersity index (PDI), zeta potential (ZP) and encapsulation efficiency (EE)). The results showed that the most adequate ratios of lipids and emulsifiers were 6.65:2.85 and 4.2:0.3 (%, w/w), with values of 84.92 nm, 0.18, −18.20 mV and 95.48% for particle size, PDI, ZP and EE, respectively. This formulation was selected for further studies related to the optimization of critical process parameters (CPPs). Full article

Background: In recent years, a growing concern about resistance to anti-infective agents has emerged1. One of the most common microbial agents is Candida albicans. Under certain conditions, C. albicans can cause infections of skin and mucosal tissues. Nystatin (Nys) is a broad-spectrum antifungal, which is indicated for the treatment of mucosal infections caused by Candida ssp such as patients under radiological treatment. Nys is a photosensitive drug and very poorly soluble in aqueous media. Therefore, microencapsulation can be the solution for its limiting factors2–3. Purpose: The aim of this work was to design, develop and characterize two types of microparticles as appropriate nystatin delivery systems for topical use: alginate microparticles (AM) and chitosan coated alginate microparticles (CCM). Methods: The formulation of the microparticles was based on the emulsification/internal gelation methodology with modification4. First, a water in oil W/O emulsion was created. Sodium alginate aqueous solution, CaCO3 and Nys were the ingredients of the internal phase, and vegetable oil the external phase. The resulting microparticles were characterized in terms of particle size, percentage yield (PY), loading capacity (LD), encapsulation efficiency (EE) and mucoadhesion ability. Results and Discussion: Microparticles ranged from 51.21 μm for AM to 57.20 μm for CCM. The PY values were 83.26% and for 79.67% AM and CCM, respectively. The LD values for the inside/surface were 6.78%/0.40% for AM and 4.87%/0.91% for CCM. The values of EE for inside and surface were 81.12%/12.07% for AM and 85.08%/9.19% for CCM. CCM was the system that exhibited the best mucoadhesive properties. Conclusions: The ability of these systems to adhere to mucous membranes has great appeal for the treatment of localized infections. Thus, these microparticulate systems could be proposed as a suitable vehicle for this kind of mucosal infections, being an alternative therapy. Full article

Ziprasidone hydrochloride is an atypical antipsychotic agent with an anti-schizophrenic activity having less solubility and less bioavailability. The nasal route acts as a promising delivery route for CNS targeting drugs due to improved bioavailability and can reduce peripheral side effects. The main aim of the study is to prepare an in-situ nasal gel of ziprasidone-β-cyclodextrin for improvement in the bioavailability of the drug. The in-situ gel was optimized using box-Behnken design. The optimized formulation was evaluated for homogeneity, viscosity, gelation temperature, mucoadhesive strength, in-vitro permeation studies. The pharmacodynamic activity of in-situ nasal gel was checked using the locomotor activity model. According to the statistical analysis of the design expert software, all the models were found significant. The pharmacodynamic investigation of the cyclodextrin mediated in situ nasal gel showed improvement in drug activity compared to a drug, drug-complex given by oral route indicating the nasal delivery of antipsychotic drug improves its effect. Full article

Impaired fracture healing impacts patients’ quality of life and imposes a financial burden on healthcare services. Up to 10% of bone fractures result in delayed/non-union fractures, for which new treatments are urgently required. However, systemic delivery of bone anabolic molecules is often sub-optimal and can lead to significant side effects. In this study, we developed ultrasound (US) responsive nano-sized vehicles in the form of perfluorocarbon nanodroplets (NDs), as a means of targeting delivery of drugs to localised tissues. We tested the hypothesis that NDs could stably encapsulate BIO (GSK-3β inhibitor), which could then be released upon US stimulation to activate Wnt signalling and induce ossification. NDs (~280 nm) were prepared from phospholipids and liquid perfluorocarbon and their stability and drug loading was studied by NTA (Nano Tracking Analysis) and HPLC. ND cytotoxicity was assessed in patient-derived bone marrow stromal cells (BMSCs) with Alamar Blue (24 h), and in vitro bioactivity of BIO-NDs was evaluated in a 3T3 Wnt-pathway reporter cell line with luciferase readout. To investigate the acoustic behaviour of NDs, 2% agarose (LM) containing NDs was injected into a bespoke bone fracture model (Sawbones) of various geometries and stimulated by US (1 MHz, 5% duty cycle, 1 MPa, 30 s), allowing the simultaneous capture of optical images and acoustic emissions. Femoral bone hole defects (1–2 mm) were made in WT-MF1 mice (age: 8–12 wks) and DiR-labelled NDs (100 µL, 109 NDs/mL, i.v.) were injected post-fracture to determine biodistribution by IVIS imaging. NDs were stable (4 and 37 °C) and retained >90% BIO until US was applied, which caused ~100% release. ND exposure up to a concentration of 109 NDs/mL showed no cytotoxicity (24 h). BIO-loaded NDs induced Wnt pathway activation in a dose dependent manner. Biodistribution of DiR-NDs in a femoral bone hole defect model in mice demonstrated increased localisation at the fracture site (~2-fold relative to that found in healthy mice or contralateral femurs at 48 h). Full article

Magnetic-fluorescent bifunctional drug delivery system which possesses magnetic targeting and fluorescent tracing capabilities effectively improves the delivery efficiency of drugs. With the in-depth study of the properties of non-spherical microparticles, it is found that the shape of the microparticles also plays a key role in drug delivery. Because of the unique shape, rod-like microparticles have exhibited great drug molecule metabolic dynamics and excellent anti-tumor effects during the process of treatment. In this study, Fe₃O₄/NaYF₄:Eu³⁺/PLGA magnetic-fluorescent bifunctional microrods were prepared via one-step electrospraying. Unlike other methods that require complicated steps or expensive equipment, one-step electrospraying is a facile and low-energy method. The prepared magnetic-fluorescent bifunctional microrods possessed uniform rod-like morphology. Compared with magnetic-fluorescent bifunctional microspheres in same volume, it was found that the microrods showed a lower water contact angle. The results of hysteresis curve and fluorescence spectrum suggested the excellent magnetic and fluorescent properties of magnetic-fluorescent bifunctional microrods. After being co-cultured with A549 cells or endothelial cells, the cell viability testing results confirmed the wonderful biocompatibility of microrods. When the drug doxorubicin was loaded by the immersion method, the microrods showed a higher drug entrapment efficiency and drug-loading capacity in comprasion with microspheres. In addition, microrods loaded with the same drug in weight showed stronger cytostatic effects after two days of co-culture with A549 cells. In summary, the magnetic-fluorescent bifunctional microrods prepared via one-step electrospraying will be promising candidates for biomedical applications in drug delivery, targeting and tracking. Full article

The creation of co-crystals as a route to creating new pharmaceutical phases with modified or defined physicochemical properties is an area of intense research. Much of the current research has focused on creating new phases for numerous active pharmaceutical ingredients (APIs) to alter physical properties such as low solubilities, enhancing processability or stability. Such studies have identified suitable co-formers and common bonding motifs to aid with the design of new co-crystals but understanding how the changes in the molecular structure of the components are reflected in the packing and resulting properties is still lacking. This lack of insight means that the design and growth of new co-crystals is still a largely empirical process with co-formers selected and then attempts to grow the different materials undertaken to evaluate the resulting properties. This work will report on the results of a combination of crystal structure database analysis with computational chemistry studies to identify what structural features are retained across a selection of families of co-crystals with common components. The competition between different potential hydrogen bonding motifs was evaluated using ab initio quantum mechanical calculations and this was related to the commonality in the packing motifs when observed. It is found while the stronger local bonding motifs are often retained within systems, the balance of weaker long-range packing forces gives rise to many subtle shifts in packing leading to greater challenges in the prediction of final crystal structures. Full article

The aim of this research work was to develop n-propyl gallate-encapsulated solid lipid nanoparticles (PG-SLNs) and load them into a hyaluronic acid (HA)-based hydrogel (HG) for intranasal delivery. Simple modified solvent injection technique was used for the preparation of the PG-SLNs via the quality-by design (QbD) approach. The optimized PG-SLNs, with an average hydrodynamic diameter of 103 ± 46.04 nm, polydispersity index (PDI) of 0.16 ± 0.001 and zeta potential of −36 ± 4.78 mV, were obtained. The percentage yield of PG-SLNs was found to be 80.78 ± 0.1%, with an encapsulation efficiency of 84 ± 0.5% and loading capacity of 60 ± 0.1%. In vitro drug release from the hydrogel-containing PG-SLNs showed sustained release profile with a lower burst effect (less than 20%) and controlled release to a greater extent within 720 min following diffusion-based release kinetics. The in vitro permeability studies showed the total permeation of PG from HG was 600 μg/cm² within 60 min, showing significant permeation of PG. Findings of this work strongly emphasize that PG-SLNs-loaded hydrogel and permeation enhancer hold significant potential to be delivered through the intranasal route. Full article

To tackle one of the biggest global health problems, the resistance of microorganisms to antibiotics, a collective effort in the search for more effective agents against bacteria was required. Peptides with antimicrobial activity have been raising much attention as a promising alternative for antibiotics. Peptaibols, for instance, are a family of antimicrobial peptides (AMPs) with great biomedical potential, in which the Peptaibolin can be highlighted. This peptide has gained relevance due to its small amino acids content, only four, and its acetyl group and a phenylalaninol residue (Phol) at the N-terminal and C-terminal, respectively. Here, we report the synthesis of Peptaibolin through Solid Phase Peptide Synthesis assisted by Microwave heating (MW-SPPS) in a pre-loaded Phe-Wang resin. Starting from a loading of 0.51 mmol/g, two syntheses were made, using two different combinations of coupling reagents. The best option was DIC/Oxima, achieving a yield of 50.0%. Proton Nuclear Magnetic Resonance (¹H-NMR) studies confirmed the peptide structure, while High Performance Liquid Chromatography (HPLC) verified the peptide purity. The peptide solubility was examined against several combinations of solvents. Peptaibolin was not soluble in water, only in organic solvents or in the combination of both. Antimicrobial testing was conducted using Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, and Pseudomonas aeruginosa. Minimum inhibitory concentration studies demonstrated the resistance of bacteria to the peptide action and the peptide instability in bacterial growth conditions. Full article

Liposomes are spherical vesicles composed of natural or artificial lipids and are used as drug carriers. However, their surface needs modification with ligands to target specific tissues. Cell-derived vesicles (CVs) are bioinspired drug carriers, as they are derived from cells by physical methods. Therefore, they preserve the topology and composition of the molecules present in the plasma membrane of the parental cell, enabling in vivo organotropism and specific drug delivery. The aim of this study is the investigation of the mechanisms by which liposomes and autologous CVs are internalized by three different breast cancer cell lines. The elucidation of the mechanism is likely to result in their optimization as drug carriers and the comprehension of their sub-cellular fate. More specifically, liposomes and CVs were produced, characterized, and loaded with the fluorescent dye FITC-dextran. Uptake experiments were performed using inhibitors of the clathrin-dependent or caveolin-dependent endocytic pathways and after incubating the cells at 4 °C, where energy dependent processes are inactivated. The results indicate that the endocytosis of CVs is active, mainly via the caveolin pathway, whereas liposomes are internalized actively by both pathways and also passively, as their uptake at 4 °C is not significantly hampered. Full article

Trans-resveratrol, due to its antioxidant property, has the potential to be successfully applied in the prevention and the treatment of neurological disorders (Parkinson’s and Alzheimer’s disease). Nevertheless, its traditional administration (intravenous, oral) and bioavailability are limited by its physical-chemical characteristics (solubility, chemical instability, sensitivity to heat, UV-light and pH). The ‘nose-to-brain’ application, as an alternate administration route, represents a way to reach the brain without the limitations of the blood-brain barrier, while the use of nano-sized drug delivery systems, like the liposomes, can overcome the developmental and therapeutic issues of the formulations. This research paper shows the application of Risk Assessment (RA), the key element of the Quality by Design mindset, in the development of a liposomal resveratrol-containing formulation with brain target and nasal administration. The study intends to demonstrate the definition of the quality target product profile, the selection of the critical factors, and the application of the RA to get a detailed view on the critical parts of the development process. On these terms, the factors with the most significant impact on the product quality among the critical material attributes (phospholipids, active pharmaceutical ingredient (API) content, cholesterol ratio, surface modification), as well as the production process (temperature, oxidation and light protection) were identified; subsequently, an RA-based liposome preparation process was described. The formulation procedures of ‘nose-to-brain’ liposomal systems loaded with drugs with many limiting factors meet several risks; however, the adaption of the QbD tools helps to focus on the aimed final product quality and achieve effective experimental designs. Full article

Nanoparticles (NPs) offer noteworthy advantages in the treatment of several diseases by prompting, among other benefits, the site-specific delivery of drugs. Ultra-small nanostructured lipid carriers (usNLCs) are no exception. These correspond to a class of NPs composed of a blend of solid and liquid lipids, the latter usually in a higher proportion, which promotes a less ordered solid lipid matrix, providing a higher drug loading capacity, drug release modulation, and improved stability in comparison with other lipid nanoparticles. Several manufacturing methods have been described for obtaining usNLCs. However, a comprehensive understanding of the process is imperative to warrant the final quality of the NPs. In the present work, the hot high pressure homogenization (HPH) method, which is characterized by easy scaling-up, simplicity and ease of use, was used for the development of highly concentrated small size NLCs. Critical process parameters (CPPs) and critical material attributes (CMAs) were evaluated to investigate the manufacturing process reproducibility, inter-batch consistency, long-term formulation stability, drug loading capacity and drug release. To gain a broader understanding of this method, multivariate analysis was applied to investigate how the physicochemical properties of the usNLC are affected by the variation in CPPs/CMAs. CPPs include HPH-time, and HPH-pressure, while CMAs, such as lipid content, are also taken into consideration. The results show that a high lipid content (15% w/w), an intermediate pressure and a short HPH time seem to be the crucial parameters to obtain both a small particle size (<100 nm) and a narrow size distribution (polydispersity index <0.2) in usNLC prepared by the hot-HPH method, without affecting zeta potential (>|30| mV). Full article

Certain challenges like the presence of highly complex structure (blood–brain barrier (BBB)), P-glycoprotein efflux, and the particular enzymatic activity stand in the way of the successful delivery of the drug moieties to the brain and make them fruitless. Many efforts have been conducted to overcome the previous. Direct delivery of drugs to the brain after the intranasal application is one of those strategies since it holds a great hope to raise the chances of drug moieties to the brain. Nanoparticles could be the potential to improve nose-to-brain drug delivery since they are able to protect the encapsulated drugs from biological and/or chemical degradation and increase their penetration across biological barriers. Based on the fact that neuroinflammation is associated with neuron death and neurodegenerative diseases like Alzheimer’s, nonsteroidal anti-inflammatory drugs (NSAIDs) might play a positive role in the disease. The present study aimed to employ the QbD approach for the first time in optimizing polymeric and lipid-based nanoparticles for the nose-to-brain delivery of Meloxicam (MEL), and to perform a comparison between the pure drug and the formulated nanosystems regarding dissolution profiles, permeability, and mucoadhesiveness. Full article

Osteoarthritis (OA) is one of the most frequent degenerative joint diseases characterized by joint pain and stiffness traditionally treated with symptomatic drugs such as oral nonsteroidal anti-inflammatory drugs (NSAIDs )and, in extreme cases, with intra-articular corticoids. However, both these drugs are not exempt from adverse effects. Curcumin (Cur) has proven its anti-inflammatory properties and its potential as an anti-osteoarthritic drug. However, its low solubility hinders its usage and limits its therapeutic efficacy. To overcome this issue, drug-in-cyclodextrin–double-loaded liposomes (DCL–DL) were developed. These liposomes contained free drug in the lipid bilayer and drug–cyclodextrin complex in the aqueous compartment. The aim of this work was to evaluate the actual effectiveness of Cur–DCL–DL formulations in the OA treatment by intra-articular treatment. For this purpose, the monoiodoacetate (MIA) model of OA pain in rats was used. A single dose of samples containing Cur as DCL–DL, conventional liposomes (SL), and empty liposomes (EL, as control) were injected once intra-articularly. Paw pressure, beam balance, and incapacitation tests were performed to evaluate OA progression at 7 and 14 days. After ending the assay, animals were sacrificed, and histological evaluation of the ankle-joint tissue was performed. Results showed that DCL–DL significantly reduced pain and ameliorated the balance and gait of rats over the 14 days, compared to SL. Histological tests showed that DCL–DL had protective properties in some aspects of OA. Full article

Osteosarcoma (OS) is a rare, aggressive bone tumor that impacts mostly children and young adults. Despite numerous therapeutic efforts, OS still presents a low patient survival rate, high metastasis, and relapse occurrence. To surpass that, polymeric micelles have been researched for the targeted co-delivery of genetic material and drugs. In this work, mixed polymeric micelles with cationic properties containing polyethyleneimine (PEI), Pluronics^® F68 and P123 were prepared. Pluronic^® F68 was activated by addition of diacrylate groups and conjugated with PEI. Pluronic^® P123 was incorporated in the formulation in a ratio of 2:1 regarding the concentration of Pluronic^® F68-PEI and Pluronic^® P123. The nanosystems were structurally characterized by FTIR and NMR spectroscopy and the morphology was assessed by TEM. Particle size, polydispersity index (PDI) and zeta potential were assessed by Dynamic and Electrophoretic Light Scattering, respectively. Small-sized, irregularly shaped F68-PEI micelles were obtained, with a PDI of 0.346 and zeta potential of 12.59 mV. Incorporating Pluronic^® P123 in the formulation lowered particle size and resulted in spherical micelles. Zeta potential decreased due the presence of Pluronic^® P123, but remained positive. These results indicate a stable, small-sized nanosystem, characteristics that suggest a capability to surpass multidrug resistance and perform active targeting towards OS. Full article

Immunotherapy is considered as a new pillar of cancer treatment. However, the application of some promising immunotherapeutic antibodies, such as antibodies against certain immune-stimulatory receptors of the TNF receptor superfamily (TNFRs), including CD40, 41BB, CD27 and anti-fibroblast growth factor-inducible 14 (anti-Fn14), are limited due to their low bioactivity. It has been previously shown that the bioactivity of such anti-TNFR antibodies could be improved by crosslinking or attachment to the plasma membrane by interaction with Fcγ receptors (FcγR). Both result in the proximity of multiple antibody-bound TNFR molecules, which allow for the activation of proinflammatory signaling pathways. In this work, we have grafted antibodies on gold nanoparticles to simulate the “activating” effect of FcγR-bound, and thus plasma membrane-presented anti-TNFR antibodies. We have developed and optimized the method for the preparation of gold nanoparticles, their functionalization with poly-ethylene glycol (PEG) linkers, and grafting of antibodies on the surface. We showed here that antibodies, including the anti-Fn14 antibody PDL192, can be successfully attached to nanoparticles without affecting antigen binding. We hypothesize that conjugation of monoclonal anti-TNFR antibodies to the inorganic nanoparticles is a promising technique to boost the efficacy of these immunotherapeutic antibodies. Full article

Solid Lipid Nanoparticles (SLN) emerged in the late 20th century as versatile nanoparticle drug delivery systems. Since then, SLNs have demonstrated to be very useful for the encapsulation of a great number of actives. In particular, the role of SLN in wound healing is pretty recent, since the major part of scientific literature on this field is concentrated during 2019 and 2020. In this pilot study, we propose the formulation of a semisolid system formed by SLN embedded in an inorganic hydrogel with demonstrated wound healing activity. The hot emulsification method was used to prepare the SLN. Subsequently, the SLN were embedded in a wound healing hydrogel composed of a clay mineral (PS9) and a natural spring water (ALI). Granulometry, pH, rheology, and Transmission Electron Microscopy (TEM) microscopy were used to characterize the formulations. Results showed that the use of natural spring water does not affect the SLN’s particle size, unlike PS9, which increased them. TEM microphotographs revealed that this increase in particle size was due to SLN coalescence in the presence of PS9. The pH of all samples was stable for 3 months. Rheology was significantly influenced by the aqueous medium; better results were obtained with ALI. In conclusion, despite the necessity of some improvements, the proposed SLN formulation would be very versatile for wound healing due to the possibility of load different actives inside the SLN together with the wound healing activity of the PS9/ALI hydrogel. Full article

The mouth can be affected by important inflammatory processes resulting from localized or systemic diseases, such as diabetes, AIDS, and leukemia, among others, which are manifested in various types of buccal sores, typically presenting pain [1]. The present work focuses on the design, formulation, and characterization of four semi-solid formulations for oral mucosa in order to symptomatically treat these painful processes. The formulations have two active pharmaceutical ingredients: triamcinolone acetonide (TA) and lidocaine hydrochloride (LIDO). The formulations also contain Orabase^® as an excipient, which is a protective, hydrophobic, and anhydrous adhesive vehicle, used to retain or facilitate the application of active pharmaceutical ingredients (APIs) to the oral mucosa. After designing the formulations, the validation of the analytical method was performed to achieve reliable analytical results. Franz-type diffusion cells were used to perform drug release studies using synthetic membrane, and permeation studies using buccal mucosa, permitting the estimation of the amount and rate of TA permeated across this mucous membrane. Further, the amount of TA retained within the tissue was estimated, as this is where it performs its anti-inflammatory activity, and showed no significant differences between the 0.05% TA + LIDO and 0.1% TA + LIDO formulations (p > 0.05). Therefore, the results demonstrate the suitability of the administration of the lowest concentration of TA tested, which achieved a similar efficacy as higher concentrations and reduced the potential systemic effects of corticoid administration. Furthermore, sublingual permeation studies were carried out to evaluate a scenario of continuous contact of the tongue with the applied formulation. The four formulations studied show pseudoplastic and thixotropic behavior, ideal for topical application. These results provide evidence for the potential of these topical formulations for the treatment of inflammatory processes in the buccal mucosa. Full article