Acta Pharmaceutica最新文献

AKR1D1, a key enzyme in the aldo-keto reductase superfamily, plays a dual role in both steroid metabolism and bile acid synthesis by catalyzing the NADPH-dependent reduction of carbon-carbon double bonds, specifically converting 3-ketosteroid hormones into 5β-steroids. Positioned at the critical intersection of steroid hormone and bile acid metabolism, AKR1D1 has the potential to profoundly influence metabolic homeostasis and drug metabolism. Despite its importance, the enzyme's therapeutic implications and role in drug metabolism remain underexplored. This study presents an optimized methodology for the cloning, expression, and purification of AKR1D1 using an Escherichia coli expression system. We identified optimal conditions for ligation and precise DNA sequencing, emphasizing the need for lower DNA concentrations and higher purity. Protein expression was evaluated in E. coli strains BL21 and Rosetta, with the highest yields achieved under extended incubation at 25 °C with controlled IPTG concentrations. Using freshly transformed cells was essential for maintaining consistent protein expression. The enzyme's activity was confirmed using a spectrofluorometric assay, demonstrating efficient reduction of testosterone to 5β-DHT. This optimized methodology facilitates the production of AKR1D1 with high specific activity, establishing a valuable platform for future research. It enables a deeper investigation into AKR1D1's contributions to drug metabolism and its therapeutic potential.

Kaempferol-3-O-rutinoside (KR) has an excellent cardioprotective effect, but its mechanism of action is not clear. Network pharmacology was used to predict the signaling pathways, whereas molecular docking was used for preliminary validation of KR binding to targets. AMI model rats with ligated left anterior descending coronary arteries were established. HE staining was used to detect pathological changes, and ELISA was used to detect the expression of TNF-α and IL-6. Network pharmacology results showed PI3K-AKT signaling pathway may be the main mechanism, and molecular docking predicted that KR could bind strongly to the PI3K and AKT. KR could significantly reduce cardiac pathological changes, decrease the level of TNF-α and IL-6, and enhance the mRNA and protein expressions of PI3K and AKT. KR ameliorates HF after AMI by enhancing the expressions of PI3K and AKT, which will be helpful in elucidating the mechanism of KR through multiple techniques.

Toxicological studies of edible plant species are important to determine the safety of their consumption. Eryngium foetidum is an edible plant used in some countries for seasoning food and as a natural remedy in folk medicine. Despite this species' gastronomic and medicinal properties, the chemical composition and toxicity have been unclear. The objective of our investigation was to determine the toxic potential of E. foetidum in the zebrafish embryo model and identify the potential compounds involved in its toxicity by electrospray ionization liquid chromatography coupled to quadrupole-time-of-flight mass spectrometry. Acute exposure of zebrafish embryos to n-hexane extract produced higher toxicity than the other extracts in a time- and concentration-dependent fashion (coagulated embryo). A 96-h median lethal concentration (LC ₅₀) of 2.63 µg mL^-1 (CI 95 % 0.58-28.5) was calculated by probit analysis. Caudal fin hypertrophy, head, yolk sac edema, caudal region, or somite malformations were observed. Secondary metabolites such as terpenes, polyphenols, and fatty acids were identified in the n-hexane extract. Also, pollutants such as diglycidyl resorcinol ether, diisopropyl adipate, and lauryl sulfate were found in the n-hexane extract. Our study revealed that chemical pollutants could be associated with the embryonic toxicity of the n-hexane extract of E. foetidum.

Acute kidney injury (AKI) is a growing global health issue with no effective treatments. This study evaluates the therapeutic effects of umbilical cord mesenchymal stem cells (UC-MSCs) on AKI caused by ischemia-reperfusion injury (IRI) in mice. Thirty mice were divided into a sham group, an IRI group, and an MSC-treated group. Renal function was assessed, and histological analysis, immunofluorescence, and real-time PCR were used to evaluate renal damage, inflammatory cell presence, and cytokine expression (TNF-α, IL-6, IL-10). Results showed that MSC treatment reduced renal damage, decreased pro-inflammatory cytokines (TNF-α, IL-6), increased anti-inflammatory IL-10, and promoted kidney repair by homing to injury sites. Thus, umbilical cord MSCs may mitigate AKI by reducing inflammation and enhancing renal repair.

The poor prognosis of glioblastoma multiforme, inadequate treatment options, and growing drug resistance urge the need to find new effective agents. Due to the significant anti-cancer potential of harmicens, hybrid compounds which comprise harmine/β-carboline and ferrocene moiety, we investigated their antiglioblastoma potential in vitro and mechanism of action (inhibition of DYRK1A, Hsp90, anti-oxidative activity). The results have shown that triazole-type harmicens, namely 5, with a ferrocene moiety in C-3 position of the β-carboline ring (IC ₅₀ = 3.7 ± 0.1 µmol L-1, SI = 12.6) and ., the C-6 substituted harmicene (IC ₅₀ = 7.4 ± 0.5 µmol L-1, SI = 5.8) exert remarkable activity and selectivity against human malignant glioblastoma cell line (U251) in vitro. On the other hand, amide-type harmicens 10, 12, and 14 exhibited strong, but non-selective activity, in the low micro-molar range. Mechanistic studies revealed that among active compounds, amide-type harmicens 12 and 14 inhibit DYRK1A and Hsp90 CTD, whereas compound 14 showed pronounced antioxidative activity. Therefore, the antiproliferative activity of harmicens might be a combination of complex molecular interactions.

Since honey has a therapeutic role in the treatment of many diseases, we investigated the content of phenolic compounds and the antioxidant activity in acacia (Robinia pseudoacacia L.), chestnut (Castanea sativa Mill.) and lime-tree (Tilia spp.) honey originating from Croatia and Germany. Total phenols, flavonols, and flavanols contents were observed at higher levels in Croatian Castanea honey compared to German Castanea honey. Significant higher values of total flavanols and hydroxycinnamic acids were measured in Croatian Tilia honey compared to German Tilia honey. For Robinia honey, significantly higher values of total phenols and flavonols were observed in almost all Croatian honey samples compared to German honey. Croatian honey samples had higher antioxidant activity compared to German honey samples with most tested methods. The highest total phenols, total flavanols, ABTS, DPPH, and FRAP values were measured in Castanea honey, then in Robinia honey, and the lowest values in Tilia honey samples. With new developed HPLC method, pinobanksin, pinocembrin, and chrysin were identified in the majority of honey samples. Our results imply that both botanical and geographical origin influence the final quality of phenolic compounds and antioxidant activity in honey. A high positive correlation between the results of antioxidant activity and polyphenols was detected.

Converting macrocycle lactams into bicyclic lactams is proposed as an additional way to further increase the metabolic stability of peptide-based drugs. Unfortunately, the synthesis of bicyclic lactams has to start almost from scratch. This study explores the Hofmann-Löffler-Freytag (HLF) reaction mechanism and products as a potential late-stage functionalisation strategy for facile conversion of macrocyclic to bicyclic ring. Laurolactam, a macrocyclic amide, exhibits significant potential for transformation into bioactive bicyclic structures with smaller, β-, γ-, δ-, and ε-lactam rings, further increasing rigidity and hydrolytic stability. With irradiation provided by a 370 nm lamp, light-induced rearrangement reaction was monitored using nuclear magnetic resonance (NMR), while involved radical intermediates were trapped using N-tert-butyl-α-phenylnitrone (PBN) spin-trap and characterised via EPR. While only two radical adduct types were identified in the electron para magnetic resonance (EPR) (C-centered radical and chlorine radical), all eight possible products are observed in the NMR. Quantum chemical calculations provide deeper insights into reaction thermodynamics and kinetics, explaining why the N-centered radical was not observed. This research highlights the feasibility of using the HLF reaction to transform macrocyclic lactams into stable bicyclic drug candidates, paving the way for new therapeutic developments.

Zeolites are a large family of minerals and the most studied is the naturally occurring clinoptilolite. They possess anti-inflammatory, antioxidant, and detoxifying properties which makes them valuable for medicinal use. Element analysis of zeolite's composition is necessary for its precise chemical characterization, and within this work development of a suspension method for the determination of manga nese, iron, and zinc by total reflection X-ray fluorescence spec-trometry (TXRF) was presented. The Box-Behnken design based on the response surface methodology was applied to determine the optimal sample preparation conditions. The significant variables such as sample amount, volume deposition, and dispersant were selected as critical variables. Based on the results obtained, sample suspensions were prepared by weighing 10 mg of the sample and adding 1 mL of 5 % Triton X-100 with 10 mL Ga as internal standard and deposition volume was set at 10 mL. The results obtained with TXRF were comparable with those obtained with the FAAS method, indicating that this technique can be used instead of the conventional methods. Using the best analytical conditions, the limits of detection for trace elements were in the range of 0.2-0.6 mg kg^-1. Trueness and precision of the results, evaluated by CRM sample analysis, were in most cases acceptable with recoveries values in the range of 104.9-111.4 % and relative standard deviations of 2-10 % (. = 6). Zeolites showed no ability to quench free radicals nor the ability to influence dietary antioxidants.

Our study aimed to assess the prevalence of fall risk-increasing drugs (FRIDs) in a sample of community-residing older patients in Croatia and its association with negative health outcomes. An observational, cross-sectional study was conducted on older patients (65+) visiting community pharmacies in three regionally different study sites in Croatia. Data were collected using a questionnaire developed for that purpose and included components of comprehensive geriatric assessment. Prevalence of FRIDs was identified using the "Screening Tool of Older Persons Prescriptions in older adults with high fall risk" (STOPPFall). In the sample of 407 participants (median age 73 (IQR 69-70) years; 63.9 % females), 79.1 % used at least one FRID. The most common drug classes were diuretics, benzodiazepines, and opioids (in 51.1 %, 38.1 %, and 17.2 % participants, respectively). More FRIDs were prescribed to the oldest old patients (85+) and participants from poorer regions of Croatia (Slavonia) (p < 0.05). Exposition to FRIDs was identified as the significant risk factor associated with falls (OR = 1.24 (1.04-1.50); p = 0.020) and higher health-care utilization (OR = 1.29 (1.10-1.51); p = 0.001). Our study highlights the need for rationalization of FRID use. To reduce the unnecessary exposure to FRIDs in older adults, health-care professionals must consider high individualization of medication schemes regarding selection, dosing, and combinations of only necessary FRIDs.