Acta Pharmaceutica最新文献

The polymeric immunoglobulin receptor (pIgR) mediates trans cytosis of IgA, a pivotal anti-inflammatory player of the mucosal immune system. Transcytosis mediated by pIgR entails protein effectors of vesicle-mediated transport involved in signal pathway activation that lead to the sorting of pIgR-IgA complexes from the basolateral to apical membrane. Each step of pIgR transport encompasses multiple targets for regulation, but the role of cholinergic system components, i.e. acetylcholine (ACh), the ligand of nicotinic (nAChR) and muscarinic (mAChR) receptors, is unclear. This study evaluated the effect of the cholinergic system on pIgR at transcriptional and protein levels. Accordingly, lipopolysaccharide (LPS)-primed Caco-2 cells were treated with nicotine (nAChR agonist) and/or mecamylamine (nAChR antagonist) or with muscarine (mAChR agonist) and/ or atropine (mAChR antagonist), and then pIgR was analysed in situ by immunofluorescence and by RT-qPCR. In general terms, cholinergic antagonists counteracted the upmodu latory outcome of both cholinergic agonists on both pIgR cellular location and mRNA levels. These findings suggest that the cholinergic system plays a key role in the regulation of epithelial immunity by modulating pIgR expression. The study provides insights into the interaction between the cholinergic system and intestinal immune mechanisms for future research in mucosal immunity and possible therapeutic strategies.

Scutellarin has a good myocardial protective effect. However, the underlying mechanism of scutellarin on cardiomyocyte pyroptosis remains unclear. In this study, we elucidated the mechanism of scutellarin to protect the injured myocardium. The molecular docking technique was used to predict the targets of scutellarin in protecting against myocardial injury. H9c2 cell pyroptosis was induced by lipopolysaccharide (LPS) and adenosine triphosphate (ATP). Then, the activities of CK and LDH were measured through a colourimetric assay. The level of cTnI was quantified by ELISA. mRNA expressions of NLRP3, cysteine-dependent aspartate-specific protease-1 (caspase-1), gasdermin D (GSDMD), interleukin-1β (IL-1β), and interleukin-18 (IL-18) were analyzed using RT-qPCR. Protein expressions of NLRP3, caspase-1, and GSDMD were detected by the immunofluorescence technique. Protein expression of NLRP3 was analysed by using Western blotting. Scutellarin had a good binding affinity with NLRP3, caspase-1, and GSDMD. Compared with LSP and ATP-treated cells, concentrations of 25, 50, and 100 µmol L-1 scutellarin reduced CK and LDH activities and the level of cTnI, decreased the mRNA expression of NLRP3, caspase-1, and GSDMD. In the mechanism study, scutellarin decreased mRNA expressions of NLRP3, caspase-1, GSDMD, IL-1β, and IL-18, and reduced the fluorescence expressions of NLRP3, caspase-1, and GSDMD. Scutellarin reduced the protein expression of NLRP3. Scutellarin inhibits myocardial cell pyroptosis induced by LPS and ATP, and the mechanism is related to the NLRP3/caspase-1/GSDMD signalling pathway.

Selumetinib (SEL) is a recently approved medication for paediatric patients who have neurofibromatosis type 1. It is the first approved therapy for this rare, debilitating, and disfiguring disease. Development of proper analytical platforms for SEL analysis in its marketed pharmaceutical formulation (Koselugo^® capsules) and blood plasma is highly warranted. Availability of such analytical tools would ensure SEL capsules' quality and effective therapy. This study introduces, for the first time, the development of two label-free and sensitive platforms for SEL quantification in capsules and human plasma. These platforms are microwave-assisted with an ultraviolet absorbance microplate reader (MW-UV) and reverse-phase high-performance liquid chromatography with a photodiode-array detector (HPLC-PDA). Both platforms employed the SEL native UV absorption as an analytical signal. The MW-UV measured the UV absorption in 96-well transparent plates at 255 nm. The HPLC-PDA involved chromatographic separation of SEL and tozasertib (TOZ), internal standard, on a C18 column both were detected at 255 nm. The optimum procedures of both platforms were established and validated following the ICH guidelines. The linearity ranges were 15-500 µg mL^-1 and 0.8-100 µg mL-1, with limits of quantification of 15.3 and 3.5 µg mL^-1, for MW-UV and HPLC-PDA, resp. Both platforms displayed high precision with relative standard deviation values ≤ 1.8 %, and high accuracy with recovery ranging from 98.3 to 102.3 %. The platforms were successfully applied to quantify SEL in bulk form, Koselugo^® capsules, and were preliminarily applied to human plasma analysis. Eco-friendliness assessment confirmed the adherence of both platforms to green analytical approaches. MW-UV and HPLC-PDA are simple and fast, enabling high-throughput analysis, thus introducing valuable tools for routine use in quality control and clinical laboratories for SEL quantification.

This study investigates the development and characterisation of niosome-based delivery systems for olanzapine, an antipsychotic drug. Niosomes were prepared using various grades of Span surfactants (Span 60, Span 40, and Span 20) in combination with cholesterol at different ratios. The formulations were characterised in terms of particle size, polydispersity index, zeta potential, and encapsulation efficiency. Results showed an inverse relationship between surfactant hydrophilic-lipophilic balance (HLB) values and niosome size, with Span 60 producing the smallest vesicles. Optimal formulations were achieved with a 1:1 ratio of surfactant to cholesterol. Span 60 niosomes exhibited the highest encapsulation efficiency (up to 81 ± 2.5 %) and the most negative zeta potential, indicating superior stability. In vitro release studies demonstrated sustained release profiles for all niosomal formulations compared to the free drug, with Span 60 formulations showing the slowest release rates. Release kinetics analysis revealed a Fickian diffusion-controlled mechanism best described by the Korsmeyer-Peppas model. These findings suggest that niosomal formulations, particularly those based on Span 60, offer a promising approach for improving olanzapine delivery, potentially enhancing its bioavailability and therapeutic efficacy in the treatment of psychiatric disorders.

This study aimed to investigate the anticancer effects and underlying mechanisms of an ethanolic extract of Lavandula stoechas L. flowers (LsHE) on colorectal cancer. The extract demon strated high phenolic content (230.31 ± 11.28 mg GAE g^-¹ dm) and strong antioxidant activity. HPLC analysis identified rosmarinic acid and quercetin as major constituents. HT-29 colorectal carcinoma cells and HEK-293 healthy kidney epithelial cells were treated with LsHE for 48 h. The concentration of LsHE required to inhibit 50 % of HT-29 cell viability was found to be 86.37 ± 3.07 µg mL^-1, whereas a higher concentration of 131.30 ± 9.33 µg mL^-1 was observed for HEK-293 cells. In HT-29 cells, flow cytometry analysis revealed increased early (9.7 %) and late (6.8 %) apoptotic populations following LsHE treatment (p < 0.0001). qRT-PCR analysis showed significant upregulation of TP53 and CASP3 compared to the untreated group (p < 0.05 and p < 0.01, resp.), while BAX expression was unexpectedly downregulated. These findings suggest that LsHE may trigger caspase-3-dependent apoptosis through a p53-mediated mechanism, potentially independent of the BAX/BCL-2 pathway. In conclusion, the present in vitro study highlights the potential of LsHE as a natural agent that still exerts some cytotoxicity toward normal epithelial cells and pro-apoptotic activity in colorectal cancer cells. Our findings provide a molecular basis for further in vivo studies to evaluate the possible therapeutic potential and mechanistic relevance of LsHE in CRC chemoprevention.

The quality of chamomile (Matricaria recutita) is largely determined by its content of essential oils and flavonoids, the main pharmacologically active constituents. In this study, the phyto-chemical profiling of 22 commercially available chamomile flower tea samples was aided by chemometrics, comparing loose teas of whole heads with tea bags containing comminuted flowers. Principal component analysis (PCA) and agglomerative hierarchical clustering (AHC), which included both essential oil and flavonoid constituents, showed that chamomile teas can be well-differentiated and categorised into two groups that are closely related to the pharmaceutical form and largely explain the influence of processing. Multivariate analyses of the phytochemical data matrix showed clear differences between loose chamomile tea and tea bags, with the former having a more consistent composition and an overall higher quality. The essential oil content varied widely (0.75-5.34 mL kg^-1), with only five loose teas exceeding the minimum content specified in the European Pharmacopoeia (≥ 4 mL kg^-1), while most tea bag samples did not fulfil this requirement. GC-MS analyses of essential oils revealed sesquiterpenes as predominant constituents, assigning all samples to the bisabolol oxide-rich chemotype. The total flavonoid content determined by UV/Vis spectrophotometry ranged from 0.17 to 0.55 %, whereas RP-HPLC/DAD analysis revealed that the levels of apigenin-7-glucoside in tea bag samples often did not meet pharmacopoeial standards. Partial least squares-discriminant analysis (PLS-DA) yielded a robust and statistically significant model, showing for the first time that the quality differences between loose teas and tea bags can be explained by at least four key components. These results highlight the utility of chemometric tools in chamomile quality assessment and emphasise the need for improved standardisation that supports the preference for whole flower teas to ensure therapeutic efficacy.

This study evaluates the antihyperlipidemic (pancreatic lipase inhibition assay), antiglycation (inhibition of bovine serum albumin glycation, BSA glycation), and antioxidant activity (ABTS, DPPH and FRAP assays) of ethanolic extracts from flowering parts of five widely distributed plant species in Croatia - Crocus heuffelianus Herb. (tepals), Nicotiana tabacum L. (petals), Malva sylvestris L. (petals), Calendula officinalis L. and Helianthus annuus L. (both sterile ligulate flowers). An in vitro-simulated system of human digestion was employed to assess the bioaccessibility of the selected phenolics and the stability of the extracts' antioxidant, hypolipi demic, and antiglycation potential following each digestion phase. The concentrations of l-ascorbic acid, individual flavonoids, and phenolic acids were determined using RP-HPLC analysis. Principal component analysis revealed significant differences in the content of bioactive compounds and their biological activity among the investigated plant species. All original extracts exhibited high antioxidant capacity (> 70 %) in at least one assay, with N. tabacum and H. annuus demonstrating the strongest anti-oxidant capacity throughout digestion. H. annuus contained the highest levels of total identified phenolic acids, total identified phenols, and total identified compounds, while N. tabacum and C. heuffelianus exhibited the highest total flavonoid content. Among individual compounds, protocatechuic acid, quercetin, and ferulic acid significantly contributed to antioxidant activity. N. tabacum had the strongest antihyperlipidemic potential in the original extracts, as well as in the most digestion phases. Strong BSA glycation inhibition (70-100 %) was observed in all plant extracts across various digestion phases, with the exception of C. heuffelianus, which exhibited mode rate inhibitory effects. These findings suggest that the analyzed flower-derived plant materials, some of which are often considered agricultural waste, can serve as sustainable and valuable resources of bioactive compounds for functional food, dietary supplements, and pharmaceutical applications.

The current research brings introductory data to phytochemical composition and biological potential of the methanolic extract derived from the aerial parts of Aaronsohnia factorovskyi. In vitro testing was conducted to evaluate its antioxidant, anti-inflammatory and antidiabetic activities. The total phenolics and total flavonoids contents of the extract were estimated as 52.46 ± 5.93 mg GAE g^-1 and 19.01 ± 2.50 mg QE g-1, resp. UPLC-ESI-MS analysis disclosed 14 chromatographic peaks corresponding to 19 putatively identified compounds, including flavonoids, sesquiterpenes, lignans, saponins and fatty acids. The antioxidant efficacy was evaluated using DPPH and phosphomolybdenum assays, as total antioxidant capacity equals to 12.31 ± 2.33 mg g^-1 and 17.40 ± 0.96 mg g^-1, resp. In vitro testing of the anti-inflammatory activity demonstrated characteristic concentrations for 50 % inhibition of cyclooxygenase enzymes of 20.85 ± 0.73 µg mL^-1 and 8.25 ± 0.29 µg mL^-1 against COX-1 and COX-2, resp. Moreover, the extract displayed strong inhibition of α-glucosidase and α-amylase enzymes with concentration for 50 % inhibition of 0.243 ± 0.009 mg mL^-1 and 0.275 ± 0.01 mg mL-1, resp. Molecular docking studies further supported these findings highlighting the strong binding of yamogenin 3-O-neohesperidoside, convallasaponin A and baicalin to α-glucosidase and α-amylase active sites, as evidenced by their high binding affinities that are comparable to that of the co-crystallized ligands. Altogether, these findings recommend A. factorovskyi as a promising source for bioactive constituents.

Hydroxypropyl methylcellulose (HPMC)-based formulations are commonly used in nasal sprays due to their gelling and viscosity-enhancing properties. However, data on in vitro studies that mimic patient use remain limited. This study investigated two commercial HPMC-based aqueous formulations and HPMC-based powder formulations to provide an understanding of the relationship between the physical properties, their performance, and the stability of nasal products. Physical properties, quality tests focused on shot mass and shot volume were assessed. The coverage area within the nasal cavity was examined at various angles of actuation (15°, 30°, 45°, and 80°) using a simulated inhalation model. The 45° spray angle exhibited the highest coverage area (%) within the nasal cavity. Devices containing liquid formulations demonstrated more reproducible shot mass and shot volume compared to dry powder preparations. These findings provide valuable insights for patients and manufacturers, leading to a better understanding of optimal usage and formulation effectiveness.

Aloe vera has a long history of medicinal use due to its diverse biological activities, including antioxidant, anti-inflammatory and antimicrobial. This study investigates the physicochemical and ADME (absorption, distribution, metabolism, excretion) properties of two primary anthraquinones from Aloe vera, aloin A and aloe-emodin. The focus of this research was to evaluate the lipophilicity, solubility, and pharmacokinetic profiles of aloin A and aloe-emodin through a combination of computational predictions, the shake-flask method, and chromatographic techniques. The optimised shake-flask method was successfully employed to determine the log P values of phyto-chemicals. Aloin A was found to be more hydrophilic than aloe-emodin, likely due to the presence of an attached β-d-glucopyranosyl unit. All RP-TLC and RP-HPLC lipophilicity indices were higher for aloe-emodin compared to aloin A, aligning with their log P values (obtained through in silico and shake-flask methods). IAM (immobili sed artificial membrane)-HPLC results suggest that unlikely partitioning in the n-octanol/water system or C18 chains, partition into phospholipids involves not only hydrophobic intermolecular recognition forces but also electrostatic interactions. The presence of a sugar moiety (β-d-glucopyranosyl unit) at the C-10 position of aloin A considerably enhanced its affinity to phospholipids compared to its affinity to alkyl chains. HSA (human serum albumin)-HPLC and AGP (α1-acid glycoprotein)-HPLC data confirmed aloe-emodin's stronger affinity to plasma proteins. The integration of computational and experimental approaches provided a detailed understanding of aloin A and aloe-emodin physicochemical and ADME properties.