RESUMO
In this study, in silico approaches are employed to investigate the binding mechanism of peptides derived from cowpea ß-vignin and HMG-CoA reductase. With the obtained information, we designed synthetic peptides to evaluate their in vitro enzyme inhibitory activity. In vitro, the total protein extract and <3 kDa fraction, at 5000 µg, support this hypothesis (95% and 90% inhibition of HMG-CoA reductase, respectively). Ile-Ala-Phe, Gln-Gly-Phe, and Gln-Asp-Phe peptides were predicted to bind to the substrate binding site of HMGCR via HMG-CoAR. In silico, it was established that the mechanism of HMG-CoA reductase inhibition largely entailed mimicking the interactions of the decalin ring of simvastatin and via H-bonding; in vitro studies corroborated the predictions, whereby the HMG-CoA reductase activity was decreased by 69%, 77%, and 78%, respectively. Our results suggest that Ile-Ala-Phe, Gln-Gly-Phe, and Gln-Asp-Phe peptides derived from cowpea ß-vignin have the potential to lower cholesterol synthesis through a statin-like regulation mechanism.
Assuntos
Hidroximetilglutaril-CoA Redutases/metabolismo , Inibidores de Hidroximetilglutaril-CoA Redutases/metabolismo , Peptídeos/metabolismo , Sequência de Aminoácidos , Animais , Sítios de Ligação , Domínio Catalítico , Meia-Vida , Ligação de Hidrogênio , Hidroximetilglutaril-CoA Redutases/química , Inibidores de Hidroximetilglutaril-CoA Redutases/química , Simulação de Acoplamento Molecular , Peptídeos/química , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Sinvastatina/química , Sinvastatina/metabolismo , Vigna/metabolismoRESUMO
Nanoparticles are promising mediators to enable nasal systemic and brain delivery of active compounds. However, the possibility of reaching therapeutically relevant levels of exogenous molecules in the body is strongly reliant on the ability of the nanoparticles to overcome biological barriers. In this work, three paradigmatic nanoformulations vehiculating the poorly soluble model drug simvastatin were addressed: (i) hybrid lecithin/chitosan nanoparticles (LCNs), (ii) polymeric poly-ε-caprolactone nanocapsules stabilized with the nonionic surfactant polysorbate 80 (PCL_P80), and (iii) polymeric poly-ε-caprolactone nanocapsules stabilized with a polysaccharide-based surfactant, i.e., sodium caproyl hyaluronate (PCL_SCH). The three nanosystems were investigated for their physicochemical and structural properties and for their impact on the biopharmaceutical aspects critical for nasal and nose-to-brain delivery: biocompatibility, drug release, mucoadhesion, and permeation across the nasal mucosa. All three nanoformulations were highly reproducible, with small particle size (â¼200 nm), narrow size distribution (polydispersity index (PI) < 0.2), and high drug encapsulation efficiency (>97%). Nanoparticle composition, surface charge, and internal structure (multilayered, core-shell or raspberry-like, as assessed by small-angle neutron scattering, SANS) were demonstrated to have an impact on both the drug-release profile and, strikingly, its behavior at the biological interface. The interaction with the mucus layer and the kinetics and extent of transport of the drug across the excised animal nasal epithelium were modulated by nanoparticle structure and surface. In fact, all of the produced nanoparticles improved simvastatin transport across the epithelial barrier of the nasal cavity as compared to a traditional formulation. Interestingly, however, the permeation enhancement was achieved via two distinct pathways: (a) enhanced mucoadhesion for hybrid LCN accompanied by fast mucosal permeation of the model drug, or (b) mucopenetration and an improved uptake and potential transport of whole PCL_P80 and PCL_SCH nanocapsules with delayed boost of permeation across the nasal mucosa. The correlation between nanoparticle structure and its biopharmaceutical properties appears to be a pivotal point for the development of novel platforms suitable for systemic and brain delivery of pharmaceutical compounds via intranasal administration.
Assuntos
Administração Intranasal/métodos , Materiais Biocompatíveis/química , Nanocápsulas/química , Sistemas de Liberação de Fármacos por Nanopartículas/química , Mucosa Nasal/efeitos dos fármacos , Sinvastatina/administração & dosagem , Sinvastatina/química , Animais , Transporte Biológico , Caproatos/química , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Quitosana/química , Liberação Controlada de Fármacos , Humanos , Ácido Hialurônico/análogos & derivados , Ácido Hialurônico/química , Lactonas/química , Lecitinas/química , Mucosa Nasal/metabolismo , Tamanho da Partícula , Polissorbatos/química , Coelhos , Solubilidade , Tensoativos/química , SuínosRESUMO
This study aims to produce and characterize alginate bilayer membranes composed of single membranes with varying cross-linking degrees to modulate simvastatin release, with potential to be used for wound-dressing. The single-layer and bilayer membranes were characterized by weight, thickness, surface pH, equilibrium-humidity, swelling degree, solubility, infrared spectroscopy (attenuated total reflectance Fourier-transform infrared), scanning electron microscopy, and water vapor transmission. Simvastatin diffusion and release rates were analyzed using Franz's cells; its indirect cytotoxicity was analyzed using human keratinocyte cells. The difference in the cross-linking degree (bottom and top layers) influenced the morphology of the membrane, and consequently its physical barrier properties. An in vitro release study demonstrated that the bilayer membrane could sustain drug-release for longer time as compared to the single-layer membrane, which could be potentially beneficial for long-term treatment of chronic wounds. A cell viability assay showed that simvastatin-loaded alginate membranes could be characterized as noncytotoxic, demonstrating their potential for use in wound-dressing applications.
Assuntos
Alginatos/química , Bicamadas Lipídicas/química , Sinvastatina/metabolismo , Bandagens , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Portadores de Fármacos/química , Liberação Controlada de Fármacos , Humanos , Concentração de Íons de Hidrogênio , Bicamadas Lipídicas/farmacologia , Sinvastatina/química , Sinvastatina/farmacologia , Solubilidade , Cicatrização/efeitos dos fármacosRESUMO
Aims: Lipid-core nanocapsules (LNCs) loaded with simvastatin (SV, SV-LNC) or lovastatin (LV, LV-LNC) were formulated for pulmonary administration. Methods: The LNC suspensions were characterized physicochemically, their stability was evaluated, and drug delivery by the pulmonary route was tested in vitro. Results: The loaded LNCs had a particle size close to 200 nm, a low polydispersity index, and a zeta potential around -20 mV. The encapsulation efficiency was high for SV (99.21 ± 0.7%) but low for LV (20.34 ± 1.2%). SV release from nanocapsules was slower than it was from SV in solution, with a monoexponential release profile, and the drug emitted and aerosol output rate was higher for SV-LNCs (1.58 µg/s) than for SV in suspension (0.54 µg/s). Conclusions: SV-LNCs had a median aerodynamic diameter of 3.51 µm and a highly respirable fraction (61.9%), indicating that nanoparticles are a suitable system for efficient delivery of simvastatin to the lung.
Assuntos
Inibidores de Hidroximetilglutaril-CoA Redutases/administração & dosagem , Lovastatina/administração & dosagem , Nanocápsulas/química , Sinvastatina/administração & dosagem , Sistemas de Liberação de Medicamentos , Liberação Controlada de Fármacos , Estabilidade de Medicamentos , Humanos , Inibidores de Hidroximetilglutaril-CoA Redutases/química , Lipídeos/química , Lovastatina/química , Nebulizadores e Vaporizadores , Tamanho da Partícula , Sinvastatina/químicaRESUMO
Skin wound healing presents a unique challenge because of its complex healing process. Herein, we developed a hydrophobic wound dressing to incorporate simvastatin, which has potential application in the treatment of ulcers and prevention of wound infection. For that matter, collagen hydrogels were grafted with dodecenylsuccinic anhydride (DDSA). The chemical modification was confirmed by FTIR and solid state 13 C-NMR spectroscopies while the ultrastructure was observed by scanning electron microscope (SEM) images. In contact angle measurements, a higher water droplet angle in DDSA-collagen gels was observed. This was consistent with the swelling assay, in which water absorption was 5.2 g/g for collagen and 1.9 g/g for DDSA-collagen. Additionally, viability and adhesion studies were performed. Cell adhesion decreased ~11% in DDSA-collagen and the number of viable cells showed a tendency to decrease as DDSA concentration increased but it was only significantly lower above concentrations of 12%. Modified gels were loaded with simvastatin showing higher adsorption capacity and lower release. Lastly, the antimicrobial and anti-inflammatory activity of DDSA-collagen materials were assessed. DDSA-collagen hydrogels, either unloaded or loaded with simvastatin showed sustained antimicrobial activity against Pseudomonas aeruginosa and Staphylococcus aureus for 72 hr probably due to the hydrophobic interaction of DDSA chains with bacterial cell walls. The antimicrobial activity was stronger against S. aureus. Collagen hydrogels also presented a prolonged antibacterial activity when they were loaded with simvastatin, confirming the antimicrobial properties of statins. Finally, it was observed that these materials can stimulate resident macrophages and promote an M2 profile which is desirable in wound healing processes.
Assuntos
Antibacterianos , Bandagens , Colágeno , Hidrogéis , Pseudomonas aeruginosa/crescimento & desenvolvimento , Sinvastatina , Staphylococcus aureus/crescimento & desenvolvimento , Succinatos , Animais , Antibacterianos/química , Antibacterianos/farmacocinética , Antibacterianos/farmacologia , Linhagem Celular , Colágeno/química , Colágeno/farmacocinética , Colágeno/farmacologia , Hidrogéis/química , Hidrogéis/farmacocinética , Hidrogéis/farmacologia , Camundongos , Sinvastatina/química , Sinvastatina/farmacocinética , Sinvastatina/farmacologia , Succinatos/química , Succinatos/farmacocinética , Succinatos/farmacologiaRESUMO
New guidelines for the limits of elemental impurities in drug products were introduced by the International Council for Harmonization in 2014. While the guidelines define a limit for each element, the complete quantification of the 24 elements included is, in fact, unnecessary. An accurate "pass/fail" test to determine whether the threshold was exceeded or not could be valuable in this context. In this study, a screening procedure using the features of high-resolution continuum source graphite furnace atomic absorption spectrometry for the evaluation of 12 elements in three different drugs was developed. The three-dimensional absorbance spectrum including time and wavelength in the vicinity of the main line of the element allows for a pass/fail decision related to the presence or absence of the element in the sample. Additionally, the bi-dimensional absorbance-wavelength spectrum defines the elements captured in the window when additional peaks are seen in the spectrum. The analysis of the selected drugs included sample digestion, the definition of pyrolysis and atomization temperatures, determination of the limit of detection and other validation parameters for each element. The evaluation of the spectra, both three- and bi-dimensional, revealed that only three elements, Cr, Ni, and Cu, were present in the samples in amounts above the LOD and therefore "fail" in the test. Nevertheless, they were quantified, and the analysis revealed that their levels were below the permitted daily exposure, which are at least 6 times higher than the LOD of the selected elements. Operating in a routine mode, the proposed method is a good option for the evaluation of elemental impurities in drug active ingredients or drug final products.
Assuntos
Grafite/química , Losartan/química , Metais Pesados/análise , Omeprazol/química , Sinvastatina/química , Espectrofotometria AtômicaRESUMO
The aim of this study was to evaluate the release of simvastatin from scaffolds composed of poly(lactic-co-glycolic) acid (PLGA) and biphasic ceramic designed for bone engineering and to assess the physico-chemical and mechanical properties of the scaffolds. Samples with 30% and 70% porosity were obtained with 0, 2, 5, and 8 wt %. of simvastatin through the solvent evaporation technique and leaching of sucrose particles. Scaffold degradation and simvastatin release were evaluated in phosphate-buffered saline. Scaffolds were analyzed by scanning electron microscopy and microtomography for two-dimensional and three-dimensional morphological characterization of the porosity, connectivity, and intrinsic permeability. The mechanical characterization was conducted based on the compressive strength and the chemical characterization by differential scanning calorimetry and energy dispersive X-ray spectroscopy. Gradual and prolonged simvastatin release from the scaffolds was observed. The release followed the Korsmeyer kinetics model with the predominance of case II transport for 30% porosity scaffolds, and anomalous behavior for the 70% porosity samples. Simvastatin release was also influenced by the slow scaffold degradation due to the strong chemical interaction between simvastatin and PLGA, as observed by differential scanning calorimetry. The scaffolds presented spherical and sucrose crystal-shaped pores that resulted in a homogenous porosity, with a predominance of open pores, ensuring interconnectivity. Simvastatin incorporation into the scaffolds and increased porosity did not influence the mechanical properties. The scaffolds presented gradual and prolonged simvastatin release, with satisfactory physico-chemical and mechanical properties. The scaffolds presented gradual and prolonged simvastatin release, with satisfactory physico-chemical and mechanical properties, a promise for applications in bone regeneration. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 2152-2164, 2019.
Assuntos
Regeneração Óssea , Cerâmica/química , Hidroxiapatitas/química , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Sinvastatina , Animais , Implantes de Medicamento/química , Implantes de Medicamento/farmacocinética , Humanos , Sinvastatina/química , Sinvastatina/farmacocinéticaRESUMO
Cell therapy (CT) can be briefly described as the use of cells or cell components in the treatment of diseases. One of the main challenges in establishing new cell types for therapy is the low survival rates of homing cells. Glycoprotein plasminogen activator inhibitor 1 (PAI-1) is a key regulator of the plasminogen activation system, and also an essential mediator of mesenchymal stem cell (MSC) post-transplant survival rate in the target tissue. It was previously observed that the survival of cells infused into the transplanted tissue increase in the presence of PAI-1 neutralizing antibodies. Simvastatin acts at several levels in the protein cascade regulating PAI-1 levels. Thus, simvastatin-induced reduction of PAI-1 levels has a therapeutic potential by modulating the main processes involved in the creation of an inhospitable environment during the process of injury (fibrosis and cell migration). In this way, simvastatin modulates process such as migration, that plays a key role in homing and engraftment of cells after cell therapy. Due to this modulatory effect, research groups proposed the use of simvastatin as an adjuvant in different cell therapy approaches. These observations allow the proposition of the potential use of simvastatin, and possibly other statins, as an adjuvant in cell therapy, due to a mechanism of action that acts in the tissue microenvironment, promoting a better efficiency of the homing and, as a consequence, an enhancement of the paracrine effects of the stem cells in the process of tissue regeneration.
Assuntos
Adjuvantes Farmacêuticos/farmacologia , Terapia Baseada em Transplante de Células e Tecidos , Inibidor 1 de Ativador de Plasminogênio/metabolismo , Sinvastatina/farmacologia , Adjuvantes Farmacêuticos/química , Sequência de Aminoácidos , Humanos , Modelos Biológicos , Inibidor 1 de Ativador de Plasminogênio/química , Sinvastatina/químicaRESUMO
The drug in a solid dosage form must undergo dissolution before it is available for absorption from the gastrointestinal tract. Liquisolid system (LS) is a technology used for increasing aqueous solubility of the drugs, which has an important role in the dissolution and absorption phenomena. However, many factors can influence the performance and success of LS. Therefore this study aimed to evaluate through a factorial design, the factors such as drug state (solution or dispersion), nonvolatile solvent and coating material that influence the increase simvastatin (BSC II drug) apparent aqueous solubility and LS flow properties. Through numerical optimization the best formulation was selected to develop a liquisolid compact (LC) and it was evaluated by dissolution tests over commercial tablets using two dissolution media. Analyzing the data, the type of nonvolatile solvent and the state of the drug (solution or dispersion) were the factors with the greatest effects on the apparent aqueous solubility response (p < 0.0001 for both). Regarding the responses that evaluated the flow properties, the type of coating material and the type of nonvolatile solvent were the factors that influenced the Carr index (p < 0.0006, p < 0.0023, respectively) and Hausner ratio (p < 0.0006, p < 0.0014, respectively), where formulations containing Kollidon® CL were more efficient than Aerosil® (which is the most commonly used coating material for LS manufacture). These results enabled us to identify which factors were most influential and to move towards the use of new excipients in the case of Kollidon® CL. In addition, allowed a wider evaluation and understanding of LS, which is considered an important technological alternative for the increase of drug solubility.
Assuntos
Anticolesterolemiantes/química , Portadores de Fármacos/química , Sinvastatina/química , Soluções/química , Solventes/química , Algoritmos , Anticolesterolemiantes/administração & dosagem , Química Farmacêutica/métodos , Sistemas de Liberação de Medicamentos/métodos , Cinética , Sinvastatina/administração & dosagem , Solubilidade , ComprimidosRESUMO
The high index of simultaneous incidence of hypertension and hypercholesterolemia in the population of many countries demands the preparation of more efficient drugs. Therefore, there is a significant area of opportunity to provide as many alternatives as possible to treat these illnesses. Taking advantage of the solubility enhancement that can be achieved when an active pharmaceutical ingredient (API) is obtained and stabilized in its amorphous state, in the present work, new drug-drug co-amorphous formulations (Simvastatin SIM- Nifedipine NIF) with enhanced solubility and stability were prepared and characterized. Results show that the co-amorphous system (molar ratio 1:1) is more soluble than the pure commercial APIs studied separately. Aqueous dissolution profiles showed increments of solubility of 3.7 and 1.7 times for SIM and NIF, correspondingly, in the co-amorphous system. The new co-amorphous formulations, monitored in time, (molar fractions 0.3, 0.5 and 0.7 of SIM) remained stable in the amorphous state for more than one year when stored at room temperature and did not show any signs of crystallization when re-heating. Inspection on the remainder of a sample after six hours of dissolution showed no recrystallization, confirming the stability of co-amorphous system. The enhanced solubility of the co-amorphous formulations makes them promising for simultaneously targeting of hypertension and hypercholesterolemia through combination therapy.