RESUMEN
The target of the current investigation was the delivery of oseltamivir phosphate (OSE) into the lung adenocarcinoma tissues by means of designing nanosized, non-toxic and biocompatible pegylated Eudragit based NPs and investigating their anticancer and antiangiogenic activity. The rationale for this strategy is to provide a novel perspective to cancer treatment with OSE loaded pegylated ERS NPs under favor of smaller particle size, biocompatible feature, cationic characteristic, examining their selective effectiveness on lung cell lines (A549 lung cancer cell line and CCD-19Lu normal cell line) and examining antiangiogenic activity by in vivo CAM analysis. For this purpose, OSE encapsulated pegylated ERS based NPs were developed and investigated for zeta potential, particle size, encapsulation efficiency, morphology, DSC, FT-IR, 1H NMR analyses. In vitro release, cytotoxicity, determination apoptotic pathways and in vivo CAM assay were carried out. Considering characterizations, NPs showed smaller particle size, cationic zeta potential, relatively higher EE%, nearly spherical shape, amorphous matrix formation and prolonged release pattern (Peppas-Sahlin and Weibull model with Fickian and non-Fickian release mechanisms). Flow cytometry was used to assess the apoptotic pathways using the Annexin V-FITC/PI staining assay, FITC Active Caspase-3 staining assay, and mitochondrial membrane potential detection tests. Activations on caspase-3 pathways made us think that OSE loaded pegylated ERS NPs triggered to apoptosis using intrinsic pathway. As regards to the in vivo studies, OSE loaded pegylated ERS based NPs demonstrated strong and moderate antiangiogenic activity for ERS-OSE 2 and ERS-OSE 3, respectively. With its cationic character, smaller particle size, relative superior EE%, homogenous amorphous polymeric matrix constitution indicated using solid state tests, prolonged release manner, highly selective to the human lung adenocarcinoma cell lines, could trigger apoptosis intrinsically and effectively, possess good in vivo antiangiogenic activity, ERS-OSE 2 formulation is chosen as a promising candidate and a potent drug delivery system to treat lung cancer.
Asunto(s)
Resinas Acrílicas/química , Inhibidores de la Angiogénesis/farmacología , Apoptosis/efectos de los fármacos , Membrana Corioalantoides/irrigación sanguínea , Portadores de Fármacos , Neoplasias Pulmonares/tratamiento farmacológico , Nanopartículas , Neovascularización Patológica , Neovascularización Fisiológica/efectos de los fármacos , Oseltamivir/farmacología , Polietilenglicoles/química , Células A549 , Animales , Embrión de Pollo , Preparaciones de Acción Retardada , Composición de Medicamentos , Liberación de Fármacos , Humanos , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , Nanotecnología , Oseltamivir/químicaRESUMEN
The current study assessed the effects of the thalidomide and palladium (II) saccharinate complex of terpyridine on the suppression of angiogenesis-mediated cell proliferation. The viability was assessed after treatment with palladium (II) complex (1.56-100 µM) and thalidomide (0.1-400 µM) alone by using ATP assay for 48 h. Palladium (II) complex was found to inhibit growth statistically significant in a dose-dependent manner in HUVECs and promoted PARP-1 cleavage through the production of ROS. On the other hand, thalidomide did not cause any significant change in cell viability. Moreover, cell death was observed to be manifested as late apoptosis due to Annexin V/SYTOX staining after palladium (II) complex treatment however, thalidomide did not demonstrate similar results. Thalidomide and palladium (II) complex also suppressed HUVEC migration and capillary-like structure tube formation in vitro in a time-dependent manner. Palladium (II) complex (5 mg/ml) treatment showed a strong antiangiogenic effect similar to positive control thalidomide (5 mg/ml) and successfully disrupted the vasculature and reduced the thickness of the vessels compared to control (agar). Furthermore, suppression of autophagy enhanced the cell death and anti-angiogenic effect of thalidomide and palladium (II) complex. We also showed that being treated with thalidomide and palladium (II) complex inhibited phosphorylation of the signaling regulators downstream of the VEGFR2. These results provide evidence for the regulation of endothelial cell functions that are relevant to angiogenesis through the suppression of the FAK/Src/Akt/ERK1/2 signaling pathway. Our results also indicate that PLC-γ1 phosphorylation leads to activation of p-Akt and p-Erk1/2 which cause stimulation on cell proliferation at lower doses. Hence, we demonstrated that palladium (II) and thalidomide can induce cell death via the Erk/Akt/PLCγ signaling pathway and that this pathway might be a novel mechanism.
Asunto(s)
Inhibidores de la Angiogénesis/farmacología , Autofagia/efectos de los fármacos , Complejos de Coordinación/farmacología , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Quinasa 1 de Adhesión Focal/metabolismo , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Neovascularización Fisiológica/efectos de los fármacos , Fosfolipasa C gamma/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Talidomida/farmacología , Familia-src Quinasas/metabolismo , Animales , Apoptosis/efectos de los fármacos , Células Cultivadas , Embrión de Pollo , Células Endoteliales de la Vena Umbilical Humana/enzimología , Humanos , Estrés Oxidativo/efectos de los fármacos , Especies de Nitrógeno Reactivo/metabolismo , Transducción de SeñalRESUMEN
Objective: The effect of polymers used in nanoparticle (NP) production on the formulation properties is one of the few studied issues. Therefore, this study aims to formulate flurbiprofen (FLB) loaded NPs with different molecular weight (Mw) poly lactic-co-glycolic acid (PLGA) and investigate the effect of Mw on NP character. One of the most important objectives is to provide a high anti-inflammatory effect with a low dose and the anti-inflammatory efficacy of the selected optimal formulation is to be determined by in vivo hen's egg test on Chorioallantoic Membrane (HET-CAM) analysis that a new, popular and in vivo animal experiment alternative method.Significance: To determine the anti-inflammatory efficacy of the optimum formulation by HET-CAM analysis. To the best of our knowledge, this is the first report on the in vivo anti-inflammatory evaluation of FLB-loaded PLGA NP using the in vivo HET-CAM assay.Methods: Blank and FLB-loaded PLGA NPs were prepared using a nanoprecipitation technique. The cell viability test for all formulation was performed with MTT in the NIH-3T3 mouse embryonic fibroblast cell line. The anti-inflammatory activity of optimum formulation (A6) was examined using the in vivo HET-CAM assay.Results: The particle sizes (PSs) of the FLB-loaded PLGA NPs were between 175 and 198 nm. The encapsulation efficiency (EE%) was a range of 82-93%. In vitro release of NPs showed extended-release up to 144 h. The release kinetics were fitted to the Peppas-Sahlin and Weibull models. The results showed that PS, PDI, EE%, and release rates of NPs were directly related to the Mw of PLGA. There is no statistically significant difference in cell viability study was observed between blank and FLB-loaded PLGA NPs. The in vivo anti-inflammatory activity results indicated that A6 coded formulation was showed significantly good anti-inflammatory potential at low dose.Conclusions: It could be concluded that FLB-loaded NPs seem to be a promising extended-release drug delivery system for oral administration with a low dose and high efficiency.
Asunto(s)
Antiinflamatorios no Esteroideos/administración & dosificación , Portadores de Fármacos/química , Flurbiprofeno/administración & dosificación , Copolímero de Ácido Poliláctico-Ácido Poliglicólico/química , Administración Oral , Animales , Antiinflamatorios no Esteroideos/farmacocinética , Bioensayo/métodos , Embrión de Pollo , Membrana Corioalantoides , Preparaciones de Acción Retardada/administración & dosificación , Preparaciones de Acción Retardada/farmacocinética , Relación Dosis-Respuesta a Droga , Composición de Medicamentos/métodos , Evaluación Preclínica de Medicamentos , Liberación de Fármacos , Flurbiprofeno/farmacocinética , Peso Molecular , Nanopartículas/químicaRESUMEN
CONTEXT: Hypericum species including Hypericum confertum Choisy, H. hircinum L., H. hyssopifolium Chaix. subsp. elongatum (Ledeb.) Woron var. microcalycinum (Boiss. & Heldr.) Boiss. and H. perforatum L. (Clusiaceae) are used as medicinal plants in Turkey. OBJECTIVE: The anti-angiogenic evaluation of Hypericum essential oils using the chick embryo chorioallantoic membrane (CAM) assay are performed with this study for the first time. MATERIALS AND METHODS: The anti-angiogenic activity of Hypericum essential oils (0.5-5.0 mg/ml) was evaluated in vivo using the CAM assay, compared to standard anti-angiogenic substances at the same concentrations, in trice replicated independent assays. GC and GC-MS analyses were carried out simultaneously to identify the chemical compositions of the Hypericum essential oils. RESULTS: The CAM treated with H. perforatum essential oil showed anti-angiogenic effect (score 0.6 ± 0.3) at 50 µg/pellet concentration, whereas other tested Hypericum essential oils showed no effect compared to the standards (e.g. suramin score 0.5 ± 0.2). Furthermore, the tested oils showed neither membrane toxicity nor irritation at the tested concentrations. The major compound of the essential oil of H. confertum was identified as germacrene D (30.2%). The major compound of the essential oils of the H. hircinum. H. hyssopifolium subsp. elongatum var. microcalycinum and H. perforatum was identified as α-pinene (88.3, 57.8, 33.3%), respectively. DISCUSSION AND CONCLUSION: Hypericum species and in particular H. perforatum essential oil may have important effect toward wound healing and various inflammations. The data obtained in this experiment suggest further investigations on various cancers due to its anti-angiogenic effects observed.
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Inhibidores de la Angiogénesis/farmacología , Membrana Corioalantoides/efectos de los fármacos , Hypericum/química , Aceites Volátiles/farmacología , Inhibidores de la Angiogénesis/administración & dosificación , Inhibidores de la Angiogénesis/aislamiento & purificación , Animales , Embrión de Pollo , Membrana Corioalantoides/irrigación sanguínea , Relación Dosis-Respuesta a Droga , Cromatografía de Gases y Espectrometría de Masas/métodos , Aceites Volátiles/administración & dosificación , Aceites Volátiles/aislamiento & purificación , Especificidad de la Especie , TurquíaRESUMEN
The essential oil from aerial parts of Ruta chalepensis L. (Sapindales: Rutaceae) was obtained by hydrodistillation, and its chemical profile was identified using gas chromatography and gas chromatography-mass spectrometry. Compounds, 2-undecanone (43.2%), 2-nonanone (27.9%), and 2-nonyl acetate (10.6%) were the major constituents of the oil. Biting deterrent activity of R. chalepensis essential oil at 10 and 50 microg/cm2, 2-undecanone at 8.5 microg/cm2, 2-nonanone at 9 microg/cm2, and 2-nonyl acetate at 9.3 microg/cm2 was similar to DEET (N, N-diethyl-meta-toluamide) at 4.8 microg/cm2, against Aedes aegypti L. Biting deterrent activity of R. chalepensis oil at 50 microg/cm2 against Anopheles quadrimaculatus Say was statistically similar to DEET at 4.8 microg/cm2, whereas the activity was lower in the other compounds tested. In cloth patch assay, R. chalepensis essential oil was effective at 187 microg/cm2, whereas 2-undecanone was effective at 108.9 microg/cm2 against Ae. aegypti. In larval bioassays, 2-undecanone showed similar toxicity whereas toxicity of R. chalepensis essential oil and 2-nonanone was higher at 24-h posttreatment at the LD50 in An. quadrimaculatus than Ae. aegypti. This study revealed that R. chalepensis essential oil and its major compounds were active biting deterrents against Ae. aegypti at higher application rates whereas only the essential oil showed activity similar to DEET against An. quadrimaculatus. 2-undecanone was the most active compound in in vivo repellency bioassay against Ae. aegypti. Chemical composition of R. chalepensis essential oil varies because of plant production and harvest practices, and the activity level of the essential oil may depend on the source of the sample.
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Aedes/efectos de los fármacos , Anopheles/efectos de los fármacos , Control de Insectos , Repelentes de Insectos/farmacología , Insecticidas/farmacología , Aceites Volátiles/farmacología , Ruta/química , Aedes/crecimiento & desarrollo , Aedes/fisiología , Animales , Anopheles/crecimiento & desarrollo , Anopheles/fisiología , Conducta Alimentaria/efectos de los fármacos , Femenino , Cromatografía de Gases y Espectrometría de Masas , Humanos , Larva/efectos de los fármacos , Larva/crecimiento & desarrollo , MasculinoRESUMEN
BACKGROUND: The inhibition of autophagy using pharmacological inhibitors such as chloroquine may be an effective strategy to overcome chemotherapy or resistance to anti-angiogenic therapy. MATERIALS AND METHODS: The cytotoxic effect of doxorubicin (0.1-1 µM), chloroquine (0.25-32 µM) and their combination were investigated by employing ATP assay in human umbilical vein endothelial cells (HUVECs). The effect of doxorubicin and chloroquine combination was also measured using tube formation assay on Matrigel. The anti-angiogenic activities of doxorubicin (2.5 µg/pellet) and chloroquine (15 µg/pellet), their combination, and standards (50 µg/pellet) were tested in vivo using the chick embryo chorioallantoic membrane (CAM) assay. RESULTS: The combination of doxorubicin and chloroquine significantly had a stronger anti-angiogenic effect than the positive control (±)-thalidomide and doxorubicin alone in the CAM assay and in vitro tube-formation assay. CONCLUSION: Chloroquine enhanced the anti-angiogenic effect of doxorubicin on CAM at the tested concentrations.
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Inhibidores de la Angiogénesis/farmacología , Antibióticos Antineoplásicos/farmacología , Autofagia/efectos de los fármacos , Cloroquina/farmacología , Doxorrubicina/farmacología , Neovascularización Patológica/prevención & control , Adenosina Trifosfato/metabolismo , Inhibidores de la Angiogénesis/uso terapéutico , Animales , Antibióticos Antineoplásicos/uso terapéutico , Proliferación Celular/efectos de los fármacos , Embrión de Pollo , Cloroquina/uso terapéutico , Membrana Corioalantoides/efectos de los fármacos , Doxorrubicina/uso terapéutico , Sinergismo Farmacológico , Quimioterapia Combinada , Células Endoteliales de la Vena Umbilical Humana , HumanosRESUMEN
In this study, the effect of Matricaria recutita L. essential oil (MEO) on the central nervous system (CNS) of mice was investigated using some behavioral methods. Chemical profiling both by GC and GC-MS analyses of the hydrodistilled essential oil of M. recutita revealed α-bisabolol oxide A (28%), α-bisabolol oxide B (17.1%), (Z)-ß-Farnesene (15.9%) and α-bisabolol (6.8%) as the main components. Changes induced by MEO (25, 50 and 100 mg/kg) and reference drug caffeine (25 mg/kg) in spontaneous locomotor activities and motor coordinations of mice were investigated by activity cage measurements and Rota-Rod tests, respectively. Open field, social interaction and elevated plus-maze tests were applied to assess the emotional state of the animals. Further, tail-suspension test was performed for evaluating the effect of MEO on depression levels of mice. As a result, at 50 and 100 mg/kg, MEO significantly increased the numbers of spontaneous locomotor activities, exhibited anxiogenic effect in the open field, elevated plus-maze and social interaction tests and decreased the immobility times of animals in tail suspension tests. The falling latencies in Rota-Rod tests did not change. This activity profile of MEO was similar to the typical psychostimulant caffeine. The exact mechanism of action underlying this stimulant-like effect should be clarified with further detailed studies.