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Background: Trypanosoma brucei is a protozoan parasite that evades the mammalian host's adaptive immune response by antigenic variation of the highly immunogenic variant surface glycoprotein (VSG). VSGs form a dense surface coat that is constantly recycled through the endosomal system. Bound antibodies are separated in the endosome from the VSG and destroyed in the lysosome. For VSGs it has been hypothesized that pH-dependent structural changes of the VSG could occur in the more acidic environment of the endosome and hence, facilitate the separation of the antibody from the VSG. Methods: We used size exclusion chromatography, where molecules are separated according to their hydrodynamic radius to see if the VSG is present as a homodimer at both pH values. To gain information about the structural integrity of the protein we used circular dichroism spectroscopy by exposing the VSG in solution to a mixture of right- and left-circularly polarized light and analysing the absorbed UV spectra. Evaluation of protein stability and molecular dynamics simulations at different pH values was performed using different computational methods. Results: We show, for an A2-type VSG, that the dimer size is only slightly larger at pH 5.2 than at pH 7.4. Moreover, the dimer was marginally more stable at lower pH due to the higher affinity (ΔG = 353.37 kcal/mol) between the monomers. Due to the larger size, the predicted epitopes were more exposed to the solvent at low pH. Moderate conformational changes (ΔRMSD = 0.35 nm) in VSG were detected between the dimers at pH 5.2 and pH 7.4 in molecular dynamics simulations, and no significant differences in the protein secondary structure were observed by circular dichroism spectroscopy. Conclusions: Thus, the dissociation of anti-VSG-antibodies in endosomes cannot be explained by changes in pH.
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Propofol is a widely used general anesthetic in clinical practice, but its use is limited by its water-insoluble nature and associated pharmacokinetic and pharmacodynamic limitations. Therefore, researchers have been searching for alternative formulations to lipid emulsion to address the remaining side effects. In this study, novel formulations for propofol and its sodium salt Na-propofolat were designed and tested using the amphiphilic cyclodextrin (CD) derivative hydroxypropyl-ß-cyclodextrin (HPßCD). The study found that spectroscopic and calorimetric measurements suggested complex formation between propofol/Na-propofolate and HPßCD, which was confirmed by the absence of an evaporation peak and different glass transition temperatures. Moreover, the formulated compounds showed no cytotoxicity and genotoxicity compared to the reference. The molecular modeling simulations based on molecular docking predicted a higher affinity for propofol/HPßCD than for Na-propofolate/HPßCD, as the former complex was more stable. This finding was further confirmed by high-performance liquid chromatography. In conclusion, the CD-based formulations of propofol and its sodium salt may be a promising option and a plausible alternative to conventional lipid emulsions.
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Cyclodextrins (CDs) are cyclic oligosaccharide structures that could be used for theranostic applications in personalized medicine. These compounds have been widely utilized not only for enhancing drug solubility, stability, and bioavailability but also for controlled and targeted delivery of small molecules. These compounds can be complexed with various biomolecules, such as peptides or proteins, via host-guest interactions. CDs are amphiphilic compounds with water-hating holes and water-absorbing surfaces. Architectures of CDs allow the drawing and preparation of CD-based polymers (CDbPs) with optimal pharmacokinetic and pharmacodynamic properties. These polymers can be cloaked with protein corona consisting of adsorbed plasma or extracellular proteins to improve nanoparticle biodistribution and half-life. Besides, CDs have become famous in applications ranging from biomedicine to environmental sciences. In this review, we emphasize ongoing research in biomedical fields using CD-based centered, pendant, and terminated polymers and their interactions with protein corona for theranostic applications. Overall, a perusal of information concerning this novel approach in biomedicine will help to implement this methodology based on host-guest interaction to improve therapeutic and diagnostic strategies.
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Ciclodextrinas , Corona de Proteínas , Ciclodextrinas/química , Medicina de Precisión , Distribución Tisular , Polímeros/química , Preparaciones Farmacéuticas , Agua , Sistemas de Liberación de MedicamentosRESUMEN
BACKGROUND: Adequate preoperative information can lessen patient anxiety. Delivering sufficient information during a personal interview, however, is time consuming, and therefore a relevant economical aspect. We investigated whether video information given to the patient before the pre-anesthetic interview has an influence on the patient's anxiety and the duration of the interview. METHOD: We randomized 302 patients undergoing different types of anesthesia. In all, 151 patients watched a short video with general information about the anticipated anesthesia procedure. Afterward, all patients had a standard pre-anesthetic interview. Patients' anxiety and satisfaction with pre-anesthesia care were assessed after the interview using a visual analogue scale. The duration of the interview was documented. Student t-test and P < 0.05 for differences between the groups. RESULTS: There was no difference in gender, age, ASA physical status, previous anesthesia experience, and the planned anesthesia procedure between the two groups. No difference in anxiety and satisfaction with pre-anesthesia care was observed. The duration of the pre-anesthetic interview was also not different between the groups. DISCUSSION: Preoperative multimedia information did not reduce anxiety or increase the patient satisfaction undergoing anesthesia. The video containing general information did not save time in the pre-anesthetic interview.
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BACKGROUND: Anesthesia has an influence on early postoperative cognitive function. This is specifically relevant in ambulatory surgery. At discharge, patients must return to their normal life and manage simple tasks. Goal was to detect influencing factors of early postoperative cognitive dysfunction after ambulatory anesthesia. METHODS: With approval of the local ethics committee, 102 individuals scheduled for ambulatory anesthesia were examined with a specific test battery. Cued and uncued reaction time, divided and selective attention were tested prior to anesthesia and at the time of discharge. Differences between the two examinations and potential influencing factors including age, premedication, type and duration of anesthesia were evaluated with the Student t-test and linear regression. P < 0.05 considered significant. RESULTS: In all, 86 individuals completed the study. Both reaction times were reduced after anesthesia compared to before. No differences were seen for divided and selective attention. Age influenced on the post-anesthesia reaction time while all other factors did not. CONCLUSION: Reaction time but not attention as more complex cognitive function is influenced by anesthesia. Age seems to be an important factor in early postoperative cognitive dysfunction.
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Aims: Delphinidin (DEL) is a plant-derived antioxidant with clinical potential to treat inflammatory pain but suffers from poor solubility and low bioavailability. The aim of the study was to develop a well-tolerated cyclodextrin (CD)-DEL complex with enhanced bioavailability and to investigate the mechanisms behind its antinociceptive effects in a preclinical model of inflammatory pain. Results: CD-DEL was highly soluble and stable in aqueous solution, and was nontoxic. Systemic administration of CD-DEL reversed mechanical and heat hyperalgesia, while its local application into the complete Freund's adjuvant (CFA)-induced inflamed paw dose-dependently reduced mechanical hyperalgesia, paw volume, formation of the lipid peroxidation product 4-hydroxy-2-nonenal (4-HNE), and tissue migration of CD68+ macrophages. CD-DEL also directly prevented 4-HNE-induced mechanical hyperalgesia, cold allodynia, and an increase in the intracellular calcium concentration into transient receptor potential ankyrin 1 expressing cells. Both 4-HNE- and CFA-induced reactive oxygen species (ROS) levels were sensitive to CD-DEL, while its capacity to scavenge superoxide anion radicals (inhibitory concentration 50 [IC50]: 70 ± 5 µM) was higher than that observed for hydroxyl radicals (IC50: 600 ± 50 µM). Finally, CD-DEL upregulated heme oxygenase 1 that was prevented by HMOX-1 siRNA in vitro. Innovation:In vivo application of DEL to treat inflammatory pain is facilitated by complexation with CD. Apart from its antioxidant effects, the CD-DEL has a unique second antioxidative mechanism involving capturing of 4-HNE into the CD cavity followed by displacement and release of the ROS scavenger DEL. Conclusion: CD-DEL has antinociceptive, antioxidative, and anti-inflammatory effects making it a promising formulation for the local treatment of inflammatory pain.
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Antocianinas/administración & dosificación , Antiinflamatorios/administración & dosificación , Hiperalgesia/tratamiento farmacológico , beta-Ciclodextrinas/química , Aldehídos/metabolismo , Animales , Antocianinas/química , Antocianinas/farmacología , Antiinflamatorios/química , Antiinflamatorios/farmacología , Calcio/metabolismo , Modelos Animales de Enfermedad , Estabilidad de Medicamentos , Adyuvante de Freund/efectos adversos , Células HEK293 , Humanos , Hiperalgesia/inducido químicamente , Hiperalgesia/metabolismo , Masculino , Ratas , Canal Catiónico TRPA1/genética , Canal Catiónico TRPA1/metabolismoRESUMEN
The interaction between the main carrier (serum albumin, SA) of endogenous and exogenous compounds in the bloodstream of different species (human, bovine, canine, rat, rabbit, and sheep) and a general anesthetic agent (propofol, PR) was investigated using an experimental technique (high-performance liquid chromatography) and computational methods (molecular docking, molecular dynamics, sequence, and phylogenetic analyses). The obtained results revealed the differences in the PR binding affinity to various homologous forms of this protein with reliable statistics (R 2 = 0.9 and p-value < 0.005), correlating with the evolutionary relationships among SAs from different species. Additionally, the protein conformational changes (root-mean-square deviation ≈ 1.0 Å) and amino acid conservation of binding sites in protein domains were detected, contributing to the SA-PR binding modes. Overall, the outcomes from this study might provide a novel methodology to assess protein-ligand interactions and to gain some interesting insights into drug pharmacokinetics and pharmacodynamics to explain its variations among different species.
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The potential protective effect of tricetinidin as novel antioxidant is investigated and compared with selected known antioxidant substances in vitro. Dihydroethidium staining was performed to detect intracellular ROS formation and the protective effect of the antioxidant substances in combination with the superoxide-inducer antimycin a (AMA). Glutathione level, mitochondrial membrane potential and HO-1 expression were analysed for further characterization of the cellular response. The cytokinesis block micronucleus test was applied to investigate the anti-genotoxic effect of the substances against insulin induced genomic damage. AMA treatment caused a significant increase in intracellular ROS formation and insulin treatment induced a significant micronucleus induction in NRK cells. Combination of the antioxidant substances with AMA or insulin protected from the oxidative stress and the micronucleus-induction. All analysed antioxidants showed comparable effects on GSH production and mitochondrial membrane potential. Only delphinidin and tricetinidin caused an increase in HO-1 expression. Tricetinidin and delphinidin might be good candidates for development as an antioxidant supplement. Further research is necessary to show possible therapeutic and preventive effects of tricetinidin and delphinidin in vivo.
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Antocianinas/farmacología , Células Epiteliales/efectos de los fármacos , Riñón/citología , Estrés Oxidativo/efectos de los fármacos , Animales , Antocianinas/química , Antioxidantes/química , Antioxidantes/farmacología , Línea Celular , Curcumina/análogos & derivados , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Glutatión/metabolismo , Hemo-Oxigenasa 1/genética , Hemo-Oxigenasa 1/metabolismo , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Pruebas de Micronúcleos , RatasRESUMEN
Volatile anesthetic-induced preconditioning (APC) has shown to have cardiac and cerebral protective properties in both pre-clinical models and clinical trials. Interestingly, accumulating evidences demonstrate that, except from some specific characters, the underlying molecular mechanisms of APC-induced protective effects in myocytes and neurons are very similar; they share several major intracellular signaling pathways, including mediating mitochondrial function, release of inflammatory cytokines and cell apoptosis. Among all the experimental results, cortical spreading depolarization is a relative newly discovered cellular mechanism of APC, which, however, just exists in central nervous system. Applying volatile anesthetic preconditioning to clinical practice seems to be a promising cardio-and neuroprotective strategy. In this review, we also summarized and discussed the results of recent clinical research of APC. Despite all the positive experimental evidences, large-scale, long-term, more precisely controlled clinical trials focusing on the perioperative use of volatile anesthetics for organ protection are still needed.
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Anestésicos por Inhalación/farmacología , Encéfalo/metabolismo , Precondicionamiento Isquémico/métodos , Miocardio/metabolismo , Daño por Reperfusión/terapia , Anestésicos por Inhalación/efectos adversos , Animales , Apoptosis , Encéfalo/irrigación sanguínea , Encéfalo/efectos de los fármacos , Vasos Coronarios/efectos de los fármacos , Humanos , Precondicionamiento Isquémico/efectos adversosRESUMEN
All-atom molecular dynamics (MD) simulations are presented on general anesthetic propofol bound to human serum albumin (HSA) due to the drug pharmacokinetics and pharmacodynamics in the circulatory system. We implemented the explicit and implicit solvation models to compare the binding affinity of propofol at the different binding sites (PR1 and PR2) in the HSA protein. Only the implicit solvation models provided the evidence in accordance with the experimental data indicating that the HSA-ligand interactions are dominanted by hydrophobic forces due to the higher drug affinity at the PR1 position with a ΔGMM-PB/SA value of -23.44kcalmol-1. Overall, this study provides important information on the accuracy of explicit and implicit solvation models to characterize the propofol interaction with different HSA binding sites.
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Anestésicos Intravenosos/química , Simulación por Computador , Propofol/química , Albúmina Sérica/química , Sitios de Unión , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Simulación de Dinámica Molecular , Solubilidad , TermodinámicaRESUMEN
The purpose of this study was to develop and implement an in silico model of indigoid-based single-electron transistor (SET) nanodevices, which consist of indigoid molecules from natural dye weakly coupled to gold electrodes that function in a Coulomb blockade regime. The electronic properties of the indigoid molecules were investigated using the optimized density-functional theory (DFT) with a continuum model. Higher electron transport characteristics were determined for Tyrian purple, consistent with experimentally derived data. Overall, these results can be used to correctly predict and emphasize the electron transport functions of organic SETs, demonstrating their potential for sustainable nanoelectronics comprising the biodegradable and biocompatible materials. In silico model and gate coupling of indigoid single-electron nano-transistors.
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BACKGROUND: Inhibitors of cyclooxygenase, which block the formation of prostaglandin (PG) E2, are the standard treatment of inflammatory pain. These drugs, however, have serious gastrointestinal, renal, and cardiovascular side effects that limit their clinical use. Cyclodextrins are neutral glucose oligomers that form a hydrophilic outer and a hydrophobic interior cavity used to carry hydrophilic substances. Methyl-ß-cyclodextrins are used currently in several drugs as enhancers and also to deliver PGs. We therefore hypothesized that randomly methylated ß-cyclodextrins (RAMEB) could be used for pain treatment. METHODS: An in silico screening for important inflammatory mediators (eg, PGE2, substance P, bradykinin, and calcitonin gene-related peptide) was performed to predict the probability of these molecules binding to RAMEB. Thereafter, a comprehensive in vitro study investigated the complexation affinity of the best target toward RAMEB or its RAMEB-fraction L (FL) using capillary electrophoresis.Wistar rats were injected intraplantarly with complete Freund's adjuvant (CFA) for 96 hours to induce inflammatory hyperalgesia. Subsequently, rats were treated intraplantarly or intravenously either with RAMEB or RAMEB FL and compared with the respective controls. Parecoxib was used as positive control. Mechanical (paw pressure threshold, PPT) and thermal (paw withdrawal latency) nociceptive thresholds were determined before injection and at the indicated time points thereafter. Paw tissue was collected after treatments, and PGE2 and PGD2 contents were measured. Analysis of variance was used for data analysis followed by appropriate post hoc comparisons. RESULTS: In silico screening indicated that PGE2, with the highest affinity, was the best candidate for RAMEB binding. Likewise, in capillary electrophoresis experiments, RAMEB had a high affinity to form inclusion complexes with the PGE2 (stability constant [K], 360 1/M; 95% confidence interval [C]: 347.58-372.42 M). Local treatment with RAMEB alleviated CFA-induced mechanical (PPT: 76.25 g; 95% CI: 56.24-96.25 g) and thermal hyperalgesia (PPT: 8.50 seconds; 95% CI: 6.76-10.23 seconds). Moreover, a systemic administration of RAMEB decreased CFA-induced mechanical (PPT: 126.66 g; 95% CI: 114.54-138.77 g) and thermal hyperalgesia (paw withdrawal latency: 11.47 seconds; 95% CI: 9.26-13.68 seconds). RAMEB FL resulted in greater in vitro PGE2-binding capacity and decreased PG content as well as hyperalgesia in vivo to a similar extent. Motor activity of the rats was not altered by RAMEB or RAMEB FL. CONCLUSIONS: Capture of PGs by cyclodextrins could be a novel and innovative tool for the treatment of inflammatory pain and bypassing some unwanted side effects of cyclooxygenase inhibitors.
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Dinoprostona/química , Dinoprostona/uso terapéutico , Inflamación/tratamiento farmacológico , Dolor/tratamiento farmacológico , beta-Ciclodextrinas/química , Animales , Simulación por Computador , Inhibidores de la Ciclooxigenasa 2/uso terapéutico , Electroforesis Capilar , Hiperalgesia/tratamiento farmacológico , Mediadores de Inflamación , Isoxazoles/uso terapéutico , Masculino , Metilación , Dolor/inducido químicamente , Dimensión del Dolor/efectos de los fármacos , Umbral del Dolor/efectos de los fármacos , Equilibrio Postural/efectos de los fármacos , Ratas , Ratas WistarRESUMEN
In this study, we investigated the ability of the general anesthetic propofol (PR) to form inclusion complexes with modified ß-cyclodextrins, including sulfobutylether-ß-cyclodextrin (SBEßCD) and hydroxypropyl-ß-cyclodextrin (HPßCD). The PR/SBEßCD and PR/HPßCD complexes were prepared and characterized, and the blood-brain barrier (BBB) permeation potential of the formulated PR was examined in vivo for the purpose of controlled drug delivery. The PR/SBEßCD complex was found to be more stable in solution with a minimal degradation constant of 0.25 h-1, a t1/2 of 2.82 h, and a Kc of 5.19 × 103 M-1 and revealed higher BBB permeability rates compared with the reference substance (PR-LIPURO) considering the calculated brain-to-blood concentration ratio (logBB) values. Additionally, the diminished PR binding affinity to SBEßCD was confirmed in molecular dynamics simulations by a maximal Gibbs free energy of binding (ΔGbind = -18.44 kcal·mol-1), indicating the more rapid PR/SBEßCD dissociation. Overall, the results demonstrated that SBEßCD has the potential to be used as a prospective candidate for drug delivery vector development to improve the pharmacokinetic and pharmacodynamic properties of general anesthetic agents at the BBB level.
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Anestésicos Intravenosos/administración & dosificación , Barrera Hematoencefálica/metabolismo , Propofol/administración & dosificación , beta-Ciclodextrinas/administración & dosificación , 2-Hidroxipropil-beta-Ciclodextrina , Anestésicos Intravenosos/química , Anestésicos Intravenosos/farmacocinética , Animales , Preparaciones de Acción Retardada/química , Sistemas de Liberación de Medicamentos , Ratones Endogámicos C57BL , Ratones Transgénicos , Simulación de Dinámica Molecular , Propofol/química , Propofol/farmacocinética , beta-Ciclodextrinas/química , beta-Ciclodextrinas/farmacocinéticaRESUMEN
In this study, we investigated the cytotoxic effects of unmodified α-cyclodextrin (α-CD) and modified cyclodextrins, including trimethyl-ß-cyclodextrin (TRIMEB) and hydroxypropyl-ß-cyclodextrin (HPßCD), on immortalized murine microvascular endothelial (cEND) cells of the blood-brain barrier (BBB). A CellTiter-Glo viability test, performed on the cEND cells showed significant differences among the different cyclodextrins. After 24 hr of incubation, TRIMEB was the most cytotoxic, and HPßCD was non-toxic. α-CD and TRIMEB exhibited greater cytotoxicity in the Dulbecco's modified Eagle's medium than in heat-inactivated human serum indicating protective properties of the human serum. The predicted dynamic toxicity profiles (Td) for α-CD and TRIMEB indicated higher cytotoxicity for these cyclodextrins compared to the reference compound (dimethylsulfoxide). Molecular dynamics simulation of cholesterol binding to the CDs suggested that not just cholesterol but phospholipids extraction might be involved in the cytotoxicity. Overall, the results demonstrate that HPßCD has the potential to be used as a candidate for drug delivery vector development and signify a correlation between the in vitro cytotoxic effect and cholesterol binding of cyclodextrins.
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Barrera Hematoencefálica/efectos de los fármacos , Ciclodextrinas/toxicidad , Células Endoteliales/efectos de los fármacos , alfa-Ciclodextrinas/toxicidad , beta-Ciclodextrinas/toxicidad , 2-Hidroxipropil-beta-Ciclodextrina , Animales , Barrera Hematoencefálica/metabolismo , Membrana Celular/metabolismo , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Colesterol/química , Colesterol/metabolismo , Simulación por Computador , Medios de Cultivo , Ciclodextrinas/química , Ciclodextrinas/metabolismo , Sistemas de Liberación de Medicamentos , Ratones , Fosfolípidos/metabolismo , alfa-Ciclodextrinas/química , alfa-Ciclodextrinas/metabolismo , beta-Ciclodextrinas/química , beta-Ciclodextrinas/metabolismoRESUMEN
The objective of the present investigation was to study the ability of sulfobutylether-ß-cyclodextrin (SBEßCD) to form an inclusion complex with sevoflurane (SEV), a volatile anesthetic with poor water solubility. The inclusion complex was prepared, characterized and its cellular toxicity and blood-brain barrier (BBB) permeation potential of the formulated SEV have also been examined for the purpose of controlled drug delivery. The SEV-SBEßCD complex was nontoxic to the primary brain microvascular endothelial (pEND) cells at a clinically relevant concentration of sevoflurane. The inclusion complex exhibited significantly higher BBB permeation profiles as compared with the reference substance (propranolol) concerning calculated apparent permeability values (Papp). In addition, SEV binding affinity to SBEßCD was confirmed by a minimal Gibbs free energy of binding (ΔGbind) value of -1.727 ± 0.042 kcal·mol-1 and an average binding constant (Kb) of 53.66 ± 9.24 mM indicating rapid drug liberation from the cyclodextrin amphiphilic cavity.
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Anestésicos por Inhalación/metabolismo , Preparaciones de Acción Retardada , Éteres Metílicos/metabolismo , beta-Ciclodextrinas/metabolismo , Anestésicos por Inhalación/química , Anestésicos por Inhalación/farmacología , Animales , Barrera Hematoencefálica/citología , Barrera Hematoencefálica/efectos de los fármacos , Barrera Hematoencefálica/metabolismo , Encéfalo/irrigación sanguínea , Encéfalo/citología , Encéfalo/efectos de los fármacos , Encéfalo/metabolismo , Permeabilidad Capilar , Supervivencia Celular/efectos de los fármacos , Composición de Medicamentos , Células Endoteliales/citología , Células Endoteliales/efectos de los fármacos , Células Endoteliales/metabolismo , Éteres Metílicos/química , Éteres Metílicos/farmacología , Ratones , Simulación del Acoplamiento Molecular , Cultivo Primario de Células , Propranolol/metabolismo , Propranolol/farmacología , Sevoflurano , beta-Ciclodextrinas/química , beta-Ciclodextrinas/farmacologíaRESUMEN
We investigated the ionization profiles for open-ring (OR) and closed-ring (CR) forms of midazolam and drug-binding modes with heptakis-(2,3,6-tri-O-methyl)-ß-cyclodextrin (trimethyl-ß-cyclodextrin; TRIMEB) using molecular modeling techniques and quantum mechanics methods. The results indicated that the total net charges for different molecular forms of midazolam tend to be cationic for OR and neutral for CR at physiological pH levels. The thermodynamic calculations demonstrated that CR is less water-soluble than OR, mainly due to the maximal solvation energy (ΔG(CR)(solv = -9.98 kcal·mol ⻹), which has a minimal ΔG(OR)(solv) of -67.01 kcal·mol⻹. A cell viability assay did not detect any signs of TRIMEB and OR/CR-TRIMEB complex toxicity on the cEND cells after 24 h of incubation in either Dulbecco's Modified Eagles Medium or in heat-inactivated human serum. The molecular docking studies identified the more flexible OR form of midazolam as being a better binder to TRIMEB with the fluorophenyl ring introduced inside the amphiphilic cavity of the host molecule. The OR binding affinity was confirmed by a minimal Gibbs free energy of binding (ΔG(bind)) value of -5.57 ± 0.02 kcal·mol⻹, an equilibrium binding constant (K(b)) of 79.89 ± 2.706 µM, and a ligand efficiency index (LE(lig)) of -0.21 ± 0.001. Our current data suggest that in order to improve the clinical applications of midazolam via its complexation with trimethyl-ß-cyclodextrin to increase drug's overall aqueous solubility, it is important to concern the different forms and ionization states of this anesthetic. All mean values are indicated with their standard deviations.
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Endotelio Vascular/patología , Midazolam/química , beta-Ciclodextrinas/química , Animales , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Endotelio Vascular/efectos de los fármacos , Humanos , Ratones , Midazolam/farmacología , Modelos Moleculares , Termodinámica , Agua/química , beta-Ciclodextrinas/farmacologíaRESUMEN
The cytochrome P450 (CYP)3A4 enzyme affects the metabolism of most drug-like substances, and its inhibition may influence drug safety. Modulation of CYP3A4 by flavonoids, such as anthocyanins, has been shown to inhibit the mutagenic activity of mammalian cells. Considering the previous investigations addressing CYP3A4 inhibition by these substances, we studied the three-dimensional quantitative structure-activity relationship (3D-QSAR) in a series of anthocyanin derivatives as CYP3A4 inhibitors. For the training dataset (n=12), comparative molecular field analysis (CoMFA) and comparative molecular similarity index analysis (CoMSIA) yielded crossvalidated and non-crossvalidated models with a q (2) of 0.795 (0.687) and r (2) of 0.962 (0.948), respectively. The models were also validated by an external test set of four compounds with r (2) of 0.821 (CoMFA) and r (2) of 0.812 (CoMSIA). The binding affinity modes associated with experimentally derived IC50 (half maximal inhibitory concentration) values were confirmed by molecular docking into the CYP3A4 active site with r (2) of 0.66. The results obtained from this study are useful for a better understanding of the effects of anthocyanin derivatives on inhibition of carcinogen activation and cellular DNA damage.
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Bilayered oromucosal film preparations (buccal films) offer a promising way to enable drug administration via the oral cavity. Adding a non-soluble or slowly eroding/dissolving backing layer to a mucoadhesive drug-loaded layer enables unidirectional drug delivery. The aim of this study was to investigate different approaches to the manufacture of bilayered films and to examine their properties by applying different characterization methods including an optimized experimental setup for the study of drug release from bilayered films. A solvent suitability study was performed screening over 15 polymers with respect to their feasibility for viscous film formation for film preparation by solvent casting method. Two methods (double-casting and pasting) were found as suitable methods for bilayered film manufacturing. Results from drug release experiments indicated that slowly eroding hypromellose backing layer films revealed the best shielding of the drug-loaded layer to enable unidirectional drug release. In summary, manufacturing of bilayered films using the described methods was feasible. Furthermore, the use of an optimized experimental setup for drug dissolution studies enabled monitoring of drug release without delays in sampling.