Release Of Contents Research Articles

Current View on Major Natural Compounds Endowed with Antibacterial and Antiviral Effects.

Nowadays, infectious diseases of bacterial and viral origins represent a serious medical problem worldwide. In fact, the development of antibiotic resistance is responsible for the emergence of bacterial strains that are refractory even to new classes of antibiotics. Furthermore, the recent COVID-19 pandemic suggests that new viruses can emerge and spread all over the world. The increase in infectious diseases depends on multiple factors, including malnutrition, massive migration of population from developing to industrialized areas, and alteration of the human microbiota. Alternative treatments to conventional antibiotics and antiviral drugs have intensively been explored. In this regard, plants and marine organisms represent an immense source of products, such as polyphenols, alkaloids, lanthipeptides, and terpenoids, which possess antibacterial and antiviral activities. Their main mechanisms of action involve modifications of bacterial cell membranes, with the formation of pores, the release of cellular content, and the inhibition of bacterial adherence to host cells, as well as of the efflux pump. Natural antivirals can interfere with viral replication and spreading, protecting the host with the enhanced production of interferon. Of note, these antivirals are not free of side effects, and their administration to humans needs more research in terms of safety. Preclinical research with natural antibacterial and antiviral compounds confirms their effects against bacteria and viruses, but there are still only a few clinical trials. Therefore, their full exploitation and more intensive clinical studies represent the next steps to be pursued in this area of medicine.

Production and Release of Proinflammatory Mediators by the Cockroach Allergen Bla g 1 via a Shared Membrane-Destabilization Mechanism.

The cockroach allergen Bla g 1 encloses an exceptionally large hydrophobic cavity, which allows it to bind and deliver unsaturated fatty acid ligands. Bla g 1-mediated delivery of naturally occurring (nMix) ligands has been shown to destabilize lipid membranes, contributing to its digestive/antiviral functions within the source organism. However, the consequences of this activity on Bla g 1 allergenicity following human exposure remain unknown. In this work, we show that Bla g 1-mediated membrane disruption can induce a proinflammatory immune response in mammalian cells via two complementary pathways. At high concentrations, the cytotoxic activity of Bla g 1 induces the release of proinflammatory cytosolic contents including damage-associated molecular patterns (DAMPs) such as heat-shock Protein-70 (HSP70) and the cytokine interleukin-1 (IL-1β). Sublytic concentrations of Bla g 1 enhanced the ability of phospholipase A2 (PLA2) to extract and hydrolyze phospholipid substrates from cellular membranes, stimulating the production of free polyunsaturated fatty acids (PUFAs) and various downstream inflammatory lipid mediators. Both of these effects are dependent on the presence of Bla g 1's natural fatty-acid (nMix) ligands with CC50 values corresponding to the concentrations required for membrane destabilization reported in previous studies. Taken together, these results suggest that mechanisms through which Bla g 1-mediated lipid delivery and membrane destabilization could directly contribute to cockroach allergic sensitization.

Formulation and Characterization of Isoconazole Loaded Invasomal Gel for Effective Antifungal Activity

The characterization of isoconazole invasomal gel formulations (IG1–IG5) aimed to evaluate their physical attributes, drug content, and drug release kinetics for topical application. All formulations exhibited transparency and smooth texture, indicating uniform drug dispersion. While IG1 had an easily pourable consistency, IG4 displayed very good consistency, suggesting viscosity differences. Despite these variations, all formulations showed good homogeneity and high drug content, with IG4 leading in drug content percentage. Spreadability and viscosity varied among formulations, influencing ease of application and adherence to the skin. Cumulative drug release profiles revealed sustained release over 12 hours, with notable differences in release rates and extents among formulations. IG4 exhibited the most sustained release, aligning with pharmacokinetic requirements for antifungal therapy. Its release profile followed a diffusion-controlled mechanism, with a high cumulative drug release percentage of 98.95 ± 0.32% over 12 hours. Antifungal activity against Candida albicans was comparable to Isoconazole, suggesting IG4’s effectiveness in treating fungal infections. Stability studies confirmed the formulation’s stability under tested conditions, supporting its potential for further development and clinical use.

Preparation and Characterization of Febuxostat Nanosuspension as Fast Dissolving Oral Film

Background: Quickly dissolved oral films are a widely accepted method of delivering drugs and help patients adhere to treatment regimens. Nanosuspensions (NS) are colloidal dispersions of drug particles with a submicron size, and their large surface area enhances the solubility and dissolution of low-water-soluble drugs. Febuxostat (FXT) is a non-purine xanthine oxidase inhibitor with a low dissolution rate that limits its absorption. Objective: To develop fast-dissolving oral films (FDOFs) containing FXT NS and convert NS into solid dosage forms to ease administration and accelerate drug release. Methods: FXT NS was prepared using Soluplus as a stabilizer and Tween80 as a co-stabilizer through an anti-solvent precipitation technique. We prepared FDOFs using a solvent casting method, utilizing hydrophilic polymers like pullulan, polyvinyl alcohol (PVA), gelatin, and plasticizers like polyethylene glycol (PEG400) and glycerin. The study assessed the film's thickness, weight, folding endurance, drug content, disintegration time, and drug release. We validated the drug's compatibility using FTIR, and conducted a crystallinity study using DSC and X-ray powder diffraction. Results: F4 was the optimized formula prepared using PVA and PEG400. In just three minutes, the F4 dissolution rate increased significantly (99.63% vs. 11.23%) compared to the FXT ordinary film. Also, it had good mechanical properties. Conclusions: FXT NS were successfully loaded into FDOFs with accepted properties.

Hyperactivation of SREBP induces pannexin-1-dependent lytic cell death

Sterol-regulatory element binding proteins (SREBPs) are a conserved transcription factor family governing lipid metabolism. When cellular cholesterol level is low, SREBP2 is transported from the endoplasmic reticulum to the Golgi apparatus where it undergoes proteolytic activation to generate a soluble N-terminal fragment, which drives the expression of lipid biosynthetic genes. Malfunctional SREBP activation is associated with various metabolic abnormalities. In this study, we find that overexpression of the active nuclear form SREBP2 (nSREBP2) causes caspase-dependent lytic cell death in various types of cells. These cells display typical pyroptotic and necrotic signatures, including plasma membrane ballooning and release of cellular contents. However, this phenotype is independent of the gasdermin family proteins or mixed lineage kinase domain-like (MLKL). Transcriptomic analysis identifies that nSREBP2 induces expression of p73, which further activates caspases. Through whole-genome CRISPR-Cas9 screening, we find that Pannexin-1 (PANX1) acts downstream of caspases to promote membrane rupture. Caspase-3 or 7 cleaves PANX1 at the C-terminal tail and increases permeability. Inhibition of the pore-forming activity of PANX1 alleviates lytic cell death. PANX1 can mediate gasdermins and MLKL-independent cell lysis during TNF-induced or chemotherapeutic reagents (doxorubicin or cisplatin)-induced cell death. Together, this study uncovers a noncanonical function of SREBPs as a potentiator of programmed cell death and suggests that PANX1 can directly promote lytic cell death independent of gasdermins and MLKL.

Calcium lactate treatment modulated fatty acid metabolism to attenuate senescence in ‘Jinfeng’ pear fruit during storage

Fatty acid metabolism not only affects cell membrane functionality but also correlates with the fruit quality and postharvest senescence. This study aims to assess the influence of calcium lactate immersion on lipid metabolism and senescence in ‘Jinfeng’ pear fruit during room temperature storage. The results demonstrated that calcium lactate effectively suppressed malondialdehyde content, respiratory rate, mass loss, and ethylene release, as well as phospholipase D (PLD), hydroperoxide lyase (HPL), lipoxygenase (LOX), lipase (LPS), alcohol acyltransferase (AAT), and alcohol dehydrogenase (ADH) activities in ‘Jinfeng’ pears. Additionally, calcium lactate resulted in an up-regulation of the gene expression levels associated with fatty acid biosynthesis, including PcACC, PcFabG, PcFabI, PcFabZ, PcKASII, PcACP, and PcFAD. Furthermore, there was an increase in the expression of genes involved in fatty acid elongation, such as PcKCS4, PcKCS6, PcKCS7, PcKCS10, PcLACS2, PcLACS4, PcLACS8, and PcCER. Conversely, it caused a down-regulation of the gene expression levels associated with fatty acid degradation, including PcLOX, PcAAT, PcADH, PcPLD, PcACD, PcECH, and PcACOX3. Simultaneously, calcium lactate treatment diminished the content of palmitic acid, while elevating the contents of linolenic acid, linoleic acid, and oleic acid. In summary, postharvest calcium lactate-immersed can efficaciously retard senescence of ‘Jinfeng’ pear fruit through regulating fatty acid lipid metabolism.

Clathrin mediates membrane fission and budding by constricting membrane pores

Membrane budding, which underlies fundamental processes like endocytosis, intracellular trafficking, and viral infection, is thought to involve membrane coat-forming proteins, including the most observed clathrin, to form Ω-shape profiles and helix-forming proteins like dynamin to constrict Ω-profiles’ pores and thus mediate fission. Challenging this fundamental concept, we report that polymerized clathrin is required for Ω-profiles’ pore closure and that clathrin around Ω-profiles’ base/pore region mediates pore constriction/closure in neuroendocrine chromaffin cells. Mathematical modeling suggests that clathrin polymerization at Ω-profiles’ base/pore region generates forces from its intrinsically curved shape to constrict/close the pore. This new fission function may exert broader impacts than clathrin’s well-known coat-forming function during clathrin (coat)-dependent endocytosis, because it underlies not only clathrin (coat)-dependent endocytosis, but also diverse endocytic modes, including ultrafast, fast, slow, bulk, and overshoot endocytosis previously considered clathrin (coat)-independent in chromaffin cells. It mediates kiss-and-run fusion (fusion pore closure) previously considered bona fide clathrin-independent, and limits the vesicular content release rate. Furthermore, analogous to results in chromaffin cells, we found that clathrin is essential for fast and slow endocytosis at hippocampal synapses where clathrin was previously considered dispensable, suggesting clathrin in mediating synaptic vesicle endocytosis and fission. These results suggest that clathrin and likely other intrinsically curved coat proteins are a new class of fission proteins underlying vesicle budding and fusion. The half-a-century concept and studies that attribute vesicle-coat contents’ function to Ω-profile formation and classify budding as coat-protein (e.g., clathrin)-dependent or -independent may need to be re-defined and re-examined by considering clathrin’s pivotal role in pore constriction/closure.

An acidic pH environment converts necroptosis to apoptosis

Cardiac ischemia results in anaerobic metabolism and lactic acid accumulation and with time, intracellular and extracellular acidosis. Ischemia and subsequent reperfusion injury (IRI) lead to various forms of programmed cell death. Necroptosis is a major form of programmed necrosis that worsens cardiac function directly and also promotes inflammation by the release of cellular contents. Potential effects of increasing acidosis on programmed cell death and their specific components have not been well studied. While apoptosis is caspase-dependent, in contrast, necroptosis is mediated by the receptor-interacting protein kinases 1 and 3 (RIPK1/3). In our study, we observed that at physiological pH = 7.4, caspase-8 inhibition did not prevent TNFα-induced cell death in mouse cardiac vascular endothelial cells (MVECs) but promoted necroptotic cell death. As expected, necroptosis was blocked by RIPK1 inhibition. However, at pH = 6.5, TNFα induced an apoptosis-like pattern which was inhibited by caspase-8 inhibition. Interestingly phosphorylation of necroptotic molecules RIPK1, RIPK3, and mixed lineage kinase domain-like protein (MLKL) was enhanced in an acidic pH environment. However, RIPK3 and MLKL phosphorylation was self-limited which may have limited their participation in necroptosis. In addition, an acidic pH promoted apoptosis-inducing factor (AIF) cleavage and nuclear translocation. AIF RNA silencing inhibited cell death, supporting the role of AIF in this cell death. In summary, our study demonstrated that the pH of the micro-environment during inflammation can bias cell death pathways by altering the function of necroptosis-related molecules and promoting AIF-mediated cell death. Further insights into the mechanisms by which an acidic cellular micro-environment influences these and perhaps other forms of regulated cell death, may lead to therapeutic strategies to attenuate IRI.

MRI-based sensing of pH-responsive content release from mesoporous silica nanoparticles

A proof of principle study toward developing a novel methodology which could be applicable for a non-invasive monitoring of the release of cargo molecules from therapeutic and diagnostic nanoparticles, as well as for possible monitoring of tissue pH variations. This was achieved by quantifying changes in longitudinal relaxation time (T1) before and after the pH-responsive release of contrast agents, for magnetic resonance imaging (MRI), from the pores of mesoporous silica nanoparticles (MSNs). The pores were filled with the FDA-approved contrast agent Gadobutrol (GdB), and its retention inside the pores ensured by covalent attachment of β-cyclodextrin monoaldehyde to hydrazine-functionalized MSN, through acidification-cleavable hydrazone linkage. The release kinetics of GdB was measured by fluorescence spectroscopy which revealed that the release of the contrast agent was enhanced at pH 5.0 in comparison to the release at pH 6.0 and 7.4. Furthermore, the changes in T1, occurring in response to the enhanced release of GdB from the pores of MSN at weakly acidic conditions, were successfully demonstrated by MRI measurements. It is envisioned that this approach using contrast agent-loaded nanoparticles before the treatment with the drug-filled analogs, could be applied in the future for tracking the locations and efficacies of nanomedicines for therapeutic cargo delivery.Graphical Pores of of β-maleimidopropionic acid hydrazide (BMPH)-functionalized mercaptopropyl-functionalized mesoporous silica nanoparticles (MPMSNs) were loaded with the contrast agent Gadobutrol and capped with β-cyclodextrin monoaldehyde. In weakly acidic environment the release of contrast agents was induced, and the enhanced release of GdB at pH 5.0 vs. pH 7.4 was evidenced by magnetic resonance imaging measurements.

Effect of Microsphere Concentration on Catechin Release from Microneedle Arrays.

In this work, microspheres were developed by cross-linking glutaraldehyde in an aqueous gelatin solution with a surfactant and solvent. A poly(vinyl alcohol) (PVA) solution was produced and combined with catechin-loaded microspheres. Different microsphere concentrations (0%, 5%, 10%, and 15%) were applied to the PVA microneedles. The moisture content, particle size, swelling, and drug release percentage of microneedles were studied using various microsphere concentrations. Fourier transform infrared and scanning electron microscopy (SEM) investigations validated the structure of gelatin microspheres as well as their decoration in microneedles. The SEM scans revealed that spherical microspheres with a wrinkled and folded morphology were created, with no physical holes visible on the surface. The gelatin microspheres generated had a mean particle size of 20-30 μm. Ex vivo release analysis indicated that microneedles containing 10% microspheres released the most catechin, with 42.9% at 12 h and 84.4% at 24 h.

Archaeosomes for Oral Drug Delivery: From Continuous Microfluidics Production to Powdered Formulations.

Archaeosomes were manufactured from natural archaeal lipids by a microfluidics-assisted single-step production method utilizing a mixture of di- and tetraether lipids extracted from Sulfolobus acidocaldarius. The primary aim of this study was to investigate the exceptional stability of archaeosomes as potential carriers for oral drug delivery, with a focus on powdered formulations. The archaeosomes were negatively charged with a size of approximately 100 nm and a low polydispersity index. To assess their suitability for oral delivery, the archaeosomes were loaded with two model drugs: calcein, a fluorescent compound, and insulin, a peptide hormone. The archaeosomes demonstrated high stability in simulated intestinal fluids, with only 5% of the encapsulated compounds being released after 24 h, regardless of the presence of degrading enzymes or extremely acidic pH values such as those found in the stomach. In a co-culture cell model system mimicking the intestinal barrier, the archaeosomes showed strong adhesion to the cell membranes, facilitating a slow release of contents. The archaeosomes were loaded with insulin in a single-step procedure achieving an encapsulation efficiency of approximately 35%. These particles have been exposed to extreme manufacturing temperatures during freeze-drying and spray-drying processes, demonstrating remarkable resilience under these harsh conditions. The fabrication of stable dry powder formulations of archaeosomes represents a promising advancement toward the development of solid dosage forms for oral delivery of biological drugs.

Role of Efferocytosis in Health and Diseases

Efferocytosis is the process of removal of apoptotic cells through phagocytosis by the specialized cells known as efferocytes. Clearance of dead cells also plays an important role in the defensive system of organisms as efferocytosis maintains homeostasis and repairing of tissues and organs. In this process, the dying cell releases signals for identification and engulfment which is further processed by macrophages. Efferocytosis prevents the secondary necrosis and release of pro-inflammatory cellular contents. This clearance process involves interplay of signaling molecules, receptors, and other mediators that ensures prompt recognitions and removal of dying cells. Dysregulation of efferocytosis has been implicated in various pathological conditions, including autoimmune diseases, chronic inflammation, and atherosclerosis. This review focuses on some common autoimmune diseases, cardiovascular diseases, respiratory disorders, and neurodegenerative disorders due to impaired efferocytosis. To describe the pathophysiology of efferocytosis in diseases more extensive studies are required.

Metallo-Glycodendrimeric Materials against Enterotoxigenic Escherichia coli.

Conjugation of carbohydrates to nanomaterials has been extensively studied and recognized as an alternative in the biomedical field. Dendrimers synthesized with mannose at the end group and with entrapped zero-valent copper/silver could be a potential candidate against bacterial proliferation. This study is aimed at investigating the bactericidal activity of metal-glycodendrimers. The Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) reaction was used to synthesize a new mannosylated dendrimer containing 12 mannopyranoside residues in the periphery. The enterotoxigenic Escherichia coli fimbriae 4 (ETEC:F4) viability, measured at 600 nm, showed the half-inhibitory concentration (IC50) of metal-free glycodendrimers (D), copper-loaded glycodendrimers (D:Cu) and silver-loaded glycodendrimers (D:Ag) closed to 4.5 × 101, 3.5 × 101 and to 1.0 × 10-2 µg/mL, respectively, and minimum inhibitory concentration (MIC) of D, D:Cu and D:Ag of 2.0, 1.5 and 1.0 × 10-4 µg/mL, respectively. The release of bacteria contents onto broth and the inhibition of ETEC:F4 biofilm formation increased with the number of metallo-glycodendrimer materials, with a special interest in silver-containing nanomaterial, which had the highest activity, suggesting that glycodendrimer-based materials interfered with bacteria-bacteria or bacteria-polystyrene interactions, with bacteria metabolism and can disrupt bacteria cell walls. Our findings identify metal-mannose-dendrimers as potent bactericidal agents and emphasize the effect of entrapped zero-valent metal against ETEC:F4.

Acetylated galactopyranosyl N-heterocyclic monocarbene complexes of Silver(I) as novel anti-proliferative agents in a rhabdomyosarcoma cell line

Herein, four silver(I) complexes bearing acetylated d-galactopyranoside-based N-heterocyclic carbene ligands were synthesized and fully characterized by elemental analysis, NMR, and X-ray photoelectron spectroscopy. All complexes were obtained with an anomeric β-configuration and as monocarbene species. In this study, we investigated the biological effects of the silver(I) complexes 2a-d on the human rhabdomyosarcoma cell line, RD. Our results show concentration-dependent effects on cell density, growth inhibition, and activation of key signaling pathways such as Akt 1/2, ERK 1/2, and p38-MAPK, indicating their potential as anticancer agents. Notably, at 35.5 µM, the complexes induced mitochondrial network disruption, as observed with 2b and 2c, whereas with 2a, this disruption was accompanied by nuclear content release. These results provide insight into the utility of carbohydrate incorporated NHC complexes of silver(I) as new agents in cancer therapy.

Abstract PO4-06-10: Targeting necroptosis in breast cancer induced by Ophiobolin A

Abstract Abstract Necroptosis or programmed necrosis is a form of cell death distinct from apoptosis. Necroptotic cells display the character of rounding of the cell, gain in cell volume, rupture of plasma membrane and release of intra-cellular contents. Although few key necroptosis regulators including receptor interacting protein kinase 3 (RIPK3) and mixed lineage kinase domain like pseudokinase (MLKL) has been identified, their role/mechanism in inducing cell death in triple-negative breast cancer (TNBC) which lack RIPK3 expression remains elusive. Ophiobolin A is a compound naturally made by fungus and potently kills cancer cells especially TNBC. Our current study investigated the cell death mechanism(s) induced by Ophiobolin A (natural product) in breast cancer cell lines. The mechanism of cell death elicited by OpA was analyzed by applying inhibitors for specific pathways including apoptosis, necroptosis, ferroptosis and paraptosis, by analyzing markers of cell death using flow cytometry and western blotting. The formation of necrosomes was analyzed with exogeneous expression of RIPK3 tagged with green fluorescence protein (GFP) in HeLa, MDA-MB-231, MDA-MB-468 cell lines which lacks endogenous RIPK3 expression and in MCF7 estrogen receptor (ER) positive cell line. OpA induced necrosome formation was identified by formation of puncta in TNBC and HeLa cells which lack RIP3 expression, however, necrosomes were not detectable in ER positive cells. Clinical relevance of RIPK3 and MLKL has been analyzed using bioinformatic tools. Effect of OpA In vivo has been analyzed using Immune compromised SCID mice. The data obtained in the present study indicates that OpA has an increased anticancer effect in TNBC compared to the estrogen receptor (ER) positive breast cancer cells. Cells having undergone EMT were shown to have more sensitivity towards OpA when compared to uninduced cell lines, which is sensitive to Necrostatin-1, an inhibitor of necroptosis. However, apoptosis, ferroptosis and paraptosis inhibitors failed to rescue cell death which clearly confirms that OpA induces necroptosis cell death pathway in EMT undergone cells. OpA treatment significantly increased puncta in TNBC compared non-TNBC cells, further confirming the necroptotic cell death in TNBC. Bioinformatic analysis revealed that RIPK3 is negatively corelated with breast cancer and has least over-all survival in patients who lack RIPK3 expression. OpA treatment showed a significant tumor regression in tumor bearing SCID mice. All together our findings identify that OpA induce necroptosis in EMT-enriched TNBC. The study further needs an in-depth analysis of molecular mechanism induced by OpA and involvement of EMT in necroptotic cell death. Citation Format: Tolulope Ojo, Santhalakshmi Ranganathan. Targeting necroptosis in breast cancer induced by Ophiobolin A [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO4-06-10.

Ammonia-enriched biogas as an alternative fuel in diesel engines: Combustion, performance and emission analysis

This study investigates the potential of ammonia as a secondary fuel in diesel engines, amidst the rising interest in alternative fuels and the growth of electric vehicles. Unlike hydrogen, with its higher flammability and safety concerns, ammonia offers a viable, sustainable fuel source. A single-cylinder diesel engine was utilized to assess engine performance and combustion characteristics using ammonia gaseous blends. Ammonia was combined with biogas to mitigate its corrosive effects and compensate for its lower heating value. Th entire best conducted with constant ammonia flow rate of 30 LPM. Diesel was mixed with biogas in compositions of 10 LPM, 20 LPM and 30 LPM, and tested under engine loads of 25%, 50%, 75%, and 100%. The blend with the highest ammonia concentration (B30B3) exhibited the largest heat release rate, highlighting a strong correlation between ammonia content and heat release levels. This is attributed to the premixed combustion phase and the accelerated flame speed associated with ammonia. An increase in ammonia content resulted in higher brake thermal efficiency and reduced brake specific fuel consumption. All ammonia-inclusive blends showed a decrease in hydrocarbon and carbon monoxide emissions, with the lowest emissions observed in the B30B3 blend. However, the high flammability of biogas in the B30B3 blend, compared to diesel, and enhanced fuel atomization, led to increased nitrogen Oxides formation. The results clearly indicate that combination of ammonia along with the biogas enhances the combustion and reduces greenhouse gas emissions. Findings of this study provides valuable insights into the use of ammonia as a sustainable alternative in diesel engines, offering a improved balance between combustion efficiency and environmental impact.

Design and characterization of glimepiride hydrotropic solid dispersion to enhance the solubility and dissolution

Background: Glimepiride lowers blood sugar levels in the body, and treats type 2 diabetes mellitus. But the main problem with the drug is its low aqueous solubility. The primary purpose of this study is to increase its solubility in an aqueous medium by using amphiphilic hydrotropic agents instead of harmful, volatile organic solvents. Methodology: A solubility study of Glimepiride was carried out using various hydrotropic agents at 10%, 20%, 30%, and 40%. In mixed hydrography, 30% of the hydrotropic agents were chosen for making blends due to their highest solubility. The blend's solubility was raised more than 50 times at fixed concentrations of urea (20%) and sodium acetate (10%) in a mixed hydrotropic solution. The solubility of Glimepiride in distilled water is 0.0038 mg/ml; in 30% urea, 49.512 ug/ml; and in 30% sodium acetate, 40.43 ug/ml. The optimized blend prepared hydrotropic solid dispersions by physical mixing and solvent evaporation. It was evaluated for drug content, FTIR, SEM, X-ray diffraction, and in vitro drug release studies. Finally, the drug release profile of the prepared tablet is compared with an already available consumer product. Result: HSD-5 showed an in-vitro drug release of 84.77±0.44 at 90 min, which is higher than the remaining formulations, and no significant change was found in drug content or drug release after 15, 30, and 45 days of stability studies. Scanning electron microscopy (SEM) showed homogenous solid dispersion, crystallinity was determined using X-ray diffraction, and FTIR showed good drug compatibility with carriers. The drug release profile of the prepared tablet was higher than that of the available consumer product. Conclusion: This study revealed that hydrotropic agents can potentially increase glimepiride's solubility and drug release. This approach can effectively enhance the solubility of poorly water-soluble drugs.

A comparison between solvent casting and electrospinning methods for the fabrication of HPMC fast-dissolving films: investigation of the physicochemical, mechanical and drug release characteristics

The fast-dissolving film is an innovative dosage form that rapidly delivers the loaded drug upon contact with saliva. Many studies have been conducted on the fabrication of fast-dissolving films using hydroxy propyl methyl cellulose (HPMC) as a common film-forming agent with solvent-casting method or sometimes with electrospinning. However, there is no comparison of these two methods on the physicochemical, mechanical, and drug-release behavior of HPMC films. This paper aims to compare bisoprolol-containing fast-dissolving films composed of HPMC and polyethylene glycol (PEG) as plasticizer constructed by solvent-casting and electrospinning methods. Given the formulation parameters (e.g., plasticizer and polymer concentrations) and process parameters (e.g., tip-to-needle distance, voltage, and feed rate), various solvent-casting films and nanofiber mats were prepared. Films were characterized in terms of disintegration time, folding endurance, thermal behavior (using DSC), crystallin structure (using XRD), drug content, and drug release. The size and morphology of nanofibers were also evaluated by SEM. Solvent-casting film composed of 6% w/v HPMC and 6% v/v PEG400, as well as nanofiber mat composed of 13% w/v HPMC and 3% w/v PEG2000 were successfully constructed. DSC and XRD analysis revealed that the crystalline structure of bisoprolol converted into the amorphous structure in the polymeric films. Nanofibers were smooth and free of beads with a mean size of 0.66 ± 0.19 μm. In terms of flexibility, the nanofiber mats outperformed the solvent-casting films. The disintegration time of the nanofiber mat was less than that of the solvent-casting film (3 s versus 30 s). Besides, nanofibers indicated faster release. The superior performance of nanofiber mats can be attributed to their nanostructure. The results showed that the characteristics of the fast-dissolving films fabricated of HPMC, including disintegration time, release rate, and flexibility could be improved through the electrospinning process.

Enzymatically Triggered Peptide-Lipid Conjugation of Designed Membrane Active Peptides for Controlled Liposomal Release.

Possibilities for controlling the release of pharmaceuticals from liposomal drug delivery systems can enhance their efficacy and reduce their side effects. Membrane-active peptides (MAPs) can be tailored to promote liposomal release when conjugated to lipid head groups using thiol-maleimide chemistry. However, the rapid oxidation of thiols hampers the optimization of such conjugation-dependent release strategies. Here, we demonstrate a de novo designed MAP modified with an enzyme-labile Cys-protection group (phenylacetamidomethyl (Phacm)) that prevents oxidation and facilitates in situ peptide lipidation. Before deprotection, the peptide lacks a defined secondary structure and does not interact with maleimide-functionalized vesicles. After deprotection of Cys using penicillin G acylase (PGA), the peptide adopts an α-helical conformation and triggers rapid release of vesicle content. Both the peptide and PGA concentrations significantly influence the conjugation process and, consequently, the release kinetics. At a PGA concentration of 5 μM the conjugation and release kinetics closely mirror those of fully reduced, unprotected peptides. We anticipate that these findings will enable further refinement of MAP conjugation and release processes, facilitating the development of sophisticated bioresponsive MAP-based liposomal drug delivery systems.

Novel Aminocoumarin Derivatives against Phytopathogenic Fungi: Design, Synthesis and Structure-Activity Relationships.

Phytopathogenic fungi is the most devastating reason for the decrease of the agricultural production and food safety. To develop new fungicidal agents for resistance concerning, a novel series of aminocoumarin derivatives were synthesized and their fungicidal activity were investigated both in vitro and in vivo. Transmission electron microscope (TEM), scanning electron microscope (SEM), RNA-Seq, 3D-QSAR and molecular docking were applied to reveal the underlying anti-fungal mechanisms. Most of the compounds exhibited significant fungicidal activity. Notably, compound 10c had a more extensive fungicidal effect than positive control. TEM indicated that compound 10c could cause abnormal morphology of cell walls, vacuoles and release of cellular contents. Transcriptional analysis data indicated that 895 and 653 out of 1548 differential expressed genes (DEGs) were up-regulated and down-regulated respectively. The Go and KEGG enrichment indicated that the coumarin derivatives could induce significant changes of succinate dehydrogenase (SDH), Acetyl-coenzyme A synthetase (ACCA) and pyruvate dehydrogenase (PDH) genes, which contributed to the disorders of glucolipid metabolism and the dysfunction of mitochondrial. The results demonstrated that aminocoumarins with schiff-base as core moieties could be the promising lead compounds for the discovery of novel fungicides.