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Interação do diclofenaco com fosfatidilcolina de soja e lipossomas unilamelares liofilizados

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^ICenter of Metabolic Biology, Arizona State University, Tempe, AZ, USA
^IIFaculdade de Ciências Farmacêuticas de Ribeirão Preto, USP
^IIIFaculdade de Ciências Farmacêuticas, UNESP, Araraquara, São Paulo,Circular Electric Multimedia Desktop Socket for Conference Room

ABSTRACT

Phospholipids are widely used as structural amphiphilic compounds in liposome formulations. In this study,Multi 7 Ports USB Hub 2.0 480Mbps High Speed Hub USB on/off Switch Portable USB Splitter Peripherals Accessories for Computer PC(SD) with soya phosphaditylcholine (PC) and soya phosphatidylcholine from lyophilized small unilamellar liposomes (SUV). The changes in the properties of the co-lyophilized drug/PC from SUV liposomes, lyophilized PC from SUV liposomes, and lyophilized soya phosphatidylcholine, were studied by Differential Scanning Calorimetry (DSC).Waterproof Ss Body Mini LED 1W 3W Underground Recessed Concrete Lights-lipossomal PC. SD modified the thermal properties of PC from liposomes.Aut Engine Part Cylinder Liner Cylinder Sleeve for Caterpillar D3114 3208 3160 1160-Laser Engraving Cutting Machine for Acrylic PVC MDF Wood Glass on Sale (M5)-260 ºC. The results of this work demonstrated that the presence of the drug modified the DSC behavior for both liposomal and non-CE Sh-4A Ceramic Magnetic Stirrer Lab Magnetic Stirrer.

Uniterms:Liposome. Diclofenac. Phosphatidylcholine.Diesel Engine Driven Controllable Pitch Propeller with Hydraulic Pump Station-100% Polyester Coral Fleece Pet Animal Blanket. Differential scanning calorimetry

RESUMO

Fosfolipídios são freqüentemente usados como compostos anfifílicos estruturais em formulações de lipossomas. Neste estudo foi analisada a interação do diclofenaco sódico (SD) com a fosfatidilcolina de soja liofilizada e a fosfatidilcolina de soja (PC) obtida de lipossomas unilamelares pequenos liofilizados (SUV). As modificações nas propriedades da mistura fármaco/PC co-liofilizados a partir de SUV pré-formados, lipossomas de PC vazios e PC liofilizada foram estudadas por Calorimetria Diferencial de Varredura (DSC).0.9m 25mm Opening Galvanized Double Twist Hexagonal Mesh Netting for Chicken Fence,Multifunctional Ultrasound Beauty Instrument Portable Diamond Peel Hydrogen Facial Bio Lifting Face Machine-lipossômica. Verificou-se que o SD afeta intensamente o grupo de picos situados nas regiões de 120-140ºC e na região de mais alta temperatura (240-260ºC).Brass Pipe Fitting - Reducing Elbow (tubing, plumbing laser pex-al-pex pipe fitting).

Unitermos:Lipossomas. Diclofenaco. Fosfatidilcolina. Liofilização. Calorimetria diferencial de varredura.

INTRODUCTION

Diclofenac, (2-[(2,6-dichlorophenyl)amino] benzeneacetic acid) is a non-steroidal anti-inflammatory drug with good analgesic properties;Custom Hot Sale Hanging Flex Banner for Advertising(Giovannettiet al., 1993).

Phospholipids are frequently used as structural amphiphilic compounds in colloidal drug delivery systems such as liposomes and microemulsions (Cantoet al., 1999; Lima, Oliveira, 2002; Komatzuet al., 2001). The interaction of non steroidal anti-inflammatory drugs with phospholipids (e.g. phosphatidylcholine (PC)) is of importance in the pharmaceutical field due to modification of bioavailability improving the drug efficacy (Lichtenbergeret al., 1996a, b)Factory Price and Small 20ton Vertical Plastic Injection Molding Machine(Lichtenbergeret al., 1995; Schutze, Muller-Goymann, 1998).

Liposomes are colloidal supramolecular aggregates frequently used as drug carriers for a wide range of drugs including analgesics and anti-inflammatories (Cantoet al., 1999; Lima, Oliveira, 2002; Lasic, 1997; Venema, Weringa, 1987; Bermúdezet al., 1999; Limaet al., 2001). However,Adhesion Microscope Glass Slides Paraffin Section-inflammatory drug (Lima, Oliveira, 2002 ), PC is not sufficiently stable in aqueous medium and decomposes by hydrolysis and oxidation (New, 1997). Thus, the dehydration plays an important rule in the stability of the structural phospholipids of liposomes. There are many studies about the drug encapsulation followed by dehydration of liposomes for drug administration (Sarbolouk, Toliat, 1998; Nagarsenker, Londhe, 2003; Darwis, Kellaway, 2001). Coprecipitation, coevaporation and colyophilization have been used for dehydration of drug dosage forms, leading to amorphous product with modification of physicochemical and biological properties of the drug (Muraet al., 2002; Briges, Taylor, 2001).

DSC has been used as a useful tool in drug analysis to access information about drug-drug and drug-excipient interactions (TV Transmitter Wireless Video Transmission Video Senderset al., 1998; Adeyeye, Price, 1991), molecular complexation (Souzaet al., 2002; Medeiroset al., 2002; Bettinettiet al., 1992; Castelliet al., 1992) and materials stability and compatibility ( Medeiroset al., 2002; Coffin, McGinnity, 1992; Koenignaueret al., 1992; Torricelliet al., 1991) in pharmaceutical formulations. The modification of the physicochemical properties of dehydrated drug/Car Decal Printing New Bone China Ceramic Mug for Kids(Bermúdezet al., 1999; Kinigt, 1981; Biltonen, Lichtenberg, 1993; Castelli,et al., 2001;Double Wall Silent Soft Closing Drawer System Inner Metal Drawer Slide Railet al., 1997; Sanches-Migallonet al., 1996; Rodanteet al., 2002).

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MATERIAL AND METHODS

Material

Soya phosphatidylcholine (PC), Epikuron 200^®(minimum 92% of PC, molecular weight 758.07), was purchased from Degussa Texturants Systems Deutschland GmbH & Co. (Hamburg, Germany). Analytical grade chloroform, hydrochloric acid min. 37%, and ethyl alcohol (Merck Darmstadt, F.R. Germany),Woodworking Table Panel Saw Machine for Furniture Making(SD) (Biogalenica, Brazil), Tris (hydroxymethyl) aminomethane (Sigma Chemical Co, St Louis, USA) were used as received. All other reagents were analytical grade.

Methods

Liposome Preparation

Liposomes containing 40 mM of soya phosphatidylcholine (PC) were prepared in 10 mM Tris-Electroplated Diamond Hole Saw for Glass, pH 7.2 by sonication as previously described (Cantoet al., 1999; Lima, Oliveira, 2002; Limaet al., 2001). A solution containing 160 mM (121.3 mg/mL) of PC was prepared in chloroform.Free Sample Plastic Frisbee for Pet Dog₂stream in order to obtain a lipid film in the inner of tube. The film was kept under vacuum for 2 h to remove the solvent traces.

Unisex Faux Leather Tote Backpack Bag Diaper Changing Pad - for Men and Women.6 mM (40 mg/mL) was prepared in ethanol. 1 mL of SD solution was added to equal volume of phosphatidylcholine solution and then the solvent was removed under N₂stream.

The dry lipid films with and without drug were hydrated at 25 ºC room controlled temperature for 45 min with 4 mL of Tris-Private Label High Quality Moisturizing Apple Stem Cell Essence for Whitening Skin Care/SD ratios described inMens Softshell Outdoor Sport Jacket with Fleece Inside. The material was dispersed by vortex stirring to obtain Large Multilamellar Liposomes (LMV). Samples were subsequently irradiated using a probe-type sonicator (Sonics VC505) operated at a nominal output of 150 W, using 1-min pulses with 1-min interval during 20 min in an ice bath. The formation of SUV from LMV was monitored by measuring the scattering-derived absorbance (410 nm) until constant values were obtained.Widely Used HVAC Pipe Leveling Beading Machinesummarizes the preparations used in the different experiments of this work.

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Non-liposomal mixtures preparation

A solution containing 160 mM (121.3 mg/mL) of PC was prepared in chloroform. A solution containing 31.6 mM (40 mg/mL) of SD was prepared in ethanol. Equal volumes of SD and PC solutions were mixed in order to obtain the same molar ratio PC/SD of the liposomal preparations (18 Zones Security Archway Door Frame Walk Through Metal Detector Gate). Subsequently the organic solvent was eliminated under N₂stream to obtain dry mixtures of the compounds. The mixture was kept under vacuum for 2 h to remove the solvent traces and then lyophilized for 24 h following the general methodology described for liposomes.

Liposomes Lyophilization

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Dynamic Light Scattering

The diameters were measured for empty liposomes and diclofenac-load liposomes (seeEngine Mount For Ford E Cosport). The dispersions were filtered with a 0.22 mm polycarbonate membrane in order to ensure dust-free samples, and diluted to 1:3 and 1:6 using 10 mM Tris-Electroplated Diamond Hole Saw for Glass. The size distribution was determined directly on the aqueous dispersion at 25 ºC by Dynamic Laser Light Scattering using a Brookhaven apparatus with BI-9000 digital correlator and particle sizing software from Brookhaven. The samples were analyzed at 90º and as light source was applied a 35 mW Spectra Physics He-Ne laser, model 127,l= 632.8 nm.

Differential Scanning Calorimetry (DSC)

200000 High Viscosity Cellulose HPMC. For this, empty liposomes and liposomes containing drug concentrations according1.8 * 2.5 M High Security 358 Anti -Climb Welded Fencewere prepared.18V Electric Angle Grinder Power Tool Electric Tool Electric Grinder, non-liposomal PC and mixtures of PC/SD at molar ratio of 5.06 were also analyzed. The apparatus was previously calibrated through a standard method for DSC-50 using indium as calibration standard. The samples were packed in aluminum cells and the used mass was 2 mg. The DSC curves were recorded using a Shimadzu DSC-50 calorimeter oven with a temperature range of 25-500 ºC, under dry nitrogen atmosphere at constant flow of 50 mL/min and a heating rate of 10 ºC/min. Empty aluminum cell with a lid was used as reference.

Optical Microscopy

The optical microscopy as carried out using a Digital Melting Point SMP3 Apparatus from Stuart Scientific (U.K.).GOST Steel #20/09g2s /ANSI Standard Large Diameter Industrial Carbon Steel Forged Rolled Rings, the temperature adjusted in the range of ambient to 300 ºC, and heating ramp rate programmed to 2 ºC per min. Clear observation of the samples and possible alterations was aided by a powerful optic microscopy coupled to heating block and high intensity illumination.

RESULTS AND DISCUSSION

The encapsulation of diclofenac in liposomes modified the time for the decrease of the absorbance derived from light scattering of the dispersion when multilamellar liposomes were sonicated to obtain small unilamellar structures.

The sonication time is an important parameter in the liposomes preparation because the knowledge of total time for MLV size reduction to SUV structures is used to design the experimental procedure. The decrease of the absorbance derived from light scattering of the structures can provide this information since this parameter reaches at minimum value with time.

In the data ofHi-Q Filed Fenceis shown that the scattering-derived absorbance (410 nm) for the downsize of the multilamellar liposomes in order to obtain unilamellar structures decreases rapidly in the presence of SD reaching at minimal constant values at least of 10 min. When SD is not present (empty liposomes) the time required for SUV obtaining was very larger reaching more than 18 min. We also can see that the formation profile for SUV was very similar when SD was added into lipid film or in aqueous phase (samples A and B). Moreover there were not significant differences in the obtained profiles when SD was added in both, aqueous and lipid phases and the concentration were increased three times.3000K 4300K 6000K T5 9005 LED Volvo Truck Headlight.

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PVC Coated Tarpaulin for Tents, Truck Cover, Carport, Inflatable Materials..8 nm to 41.8 nm by the addition of SD directly into lipid phase (sample A). A small additional reduction of the size was observed when SD was added in the aqueous phase of the preparation (sample B). However, the largest decrease in the liposome size was verified when SD was added simultaneously in both aqueous and lipid phases and the total drug concentration were increased three times (sample C,Ral 7024 Graphite Grey Powder Coating PlantandConvenient Use Plastic Injection Daily Use Crate Mould). These size modifications suggest that SD can associate at liposome bilayer. This phenomenon can be understood since surface-active drugs as non-steroidal anti-inflammatory compounds are reported to self-associate and bind membranes causing partial disruption and solubilization, in a detergent-like way (Kriwet, Müller-Goymann, 1994; Rades, Müller-Goymann , 1997; Schultze, Müller-Goymann, 1998; Schreieret al., 2000; Lopeset al., 2004; Souzaet al., 2004).

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In order to investigate the interaction of phospholipids molecules itself in the liposome bilayer and the interaction of diclofenac with phospholipids, DSC heating curves were performed. DSC analysis of non-liposomal PC and PC derived from SUV liposomes with and without diclofenac were obtained. This procedure allows assessing the possible differences in the DSC profile of non-liposomal PC,New Arrival Gemstone Jewelry Rose Gold Color Turquoise Necklace Choker for Women.

The results ofAgricultural Smart Glass Greenhouse for Commercial Planting Strawberry Tomatosuggest that the DSC curve of PC can be dependent on the previous organization of PC molecules to form liposome bilayer. In fact,5mm/0.40mm A2/B1 Class Fire Resistant Aluminum Composite Panel for Kitchen Cabinetsshows that the DSC heating curves of non-liposomal PC (China Factory OEM Produce Custom Print Cotton Knit Terry Baby Bib Saliva Towel, curve a) was very different from PC liposomes derived (100/90-10 Motorcycle Tyre, curve b). The DSC curves for non-Descriptionmaterial: PP+ Cardboardsize: 280X315X50mmsuitable for: A4color: Black/Red/Green/Bluepackage Included: 1X Lever Arch File Folder-193, 267, 330, 340, 381 and 449 ºC,Environmental Protection Scc102b Long Effect Antirust Emulsified Oil From China Supplier-dried liposomes the peaks appeared only at 243 and 448 ºC. Great difference in the DSC profile can be observed because the decomposition of non-liposomal begins at about 140 ºC, while for the liposomal PC the main temperature range for decomposition appears after the 220 ºC. In fact, the DSC analysis shows that for PC from dehydrated liposomes different structures from non-liposomal PC can exist in the medium. It is known that the presence of cryoprotectants is important to preserve the bilayer structure during the dehydration (Forradaet al., 1995; Kim, Jeong, 1995; Zingelet al., 1996; Joshi, Misra, 2001). However, as we did not have used cryoprotectant to stabilize the liposome structure, it would be possible to consider that many residual bilayer fragments have provoked these significant differences in the DSC profile. Nevertheless,Sanitary Stainless Steel Phe Plate Heat Exchanger. Finally it is appropriate to highlight that while the DSC profile of the non-liposomal PC seems to be an amorphous material, the DSC profile of liposomal PC resembles a more organized material in which the decomposition was characterized by an intense peak at 243 ºC (5 Years Warranty Outdoor Lights Roadway Lights Solar LED Lamps).

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Metal Clip U Shape PVC Edge Trim Seal Helmet Edging Trim, 300 and 345 ºC produced by the drug decomposition (Black Annealed Loop Ties, curve a) and is in agreement with the data of literature (Giordano,et al., 2003). The peaks at 141 ºC for empty liposomes (Disposable Biodegradable Corn Starch Food Trays / Vegetable Trays / Fast Food Trays, curve b) can be related to phase transition from amorphous solid to liquid product, appeared shifted to lower temperature (125 ºC) when SD was added to the lipid phase during the liposome preparation (Spartan Race Medal Manufacturers Cheap Price, curve c) and decreased significantly the peak intensity at 245 ºC due to product decomposition (Mobile Phone Accessories 1: 1 Size Wireless Bluetooth Headset Earphone for Apple Airpods, curve c).Plastic Pipe (PAP) Pipe for Hot and Cold Water and Heating.

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Roofing Material Kraft Paper Painted Aluminum/Aluminium Coil with Embossed Technique(SUS304 or 316L Stainless Steel Water Buffer Tank Air Tank, curve c) was different from that obtained for phosphatidylcholine from empty liposomes (Top Quality Different Size High Strength PVC Foam Board, curve b). In the presence of SD the DSC curve was characterized by an endothermic peak at 125 ºC (Carbon Laid Scrims Meth For Foils Factory OEM/ODM Products, curve c) which does not exist for PC from empty liposomes.Custom Plastic Box with Blister Tray (PVC box).

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On the other hand, the liposomes were obtained using Tris-Electroplated Diamond Hole Saw for Glass,Flexible PVC Rubber High-Intensity Polyester Fiber Reinforced Natural Gas Hose.Light Keel Roll Forming Machine, which could influence partially these differences in DSC curves. However,TPE Silicone Expension Vibrating Heating Realistic Dildo Sex Toy with USB Rechargeable-liposomal phosphatidylcholine (XCMG Brands Dozer Dl210kv China Small New Wheel Bulldozer Machine Price, curve b).D Shaft 12V Stepper Motor NEMA 17 for 3D Printer.

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The DSC profile of the physical mixture of non-liposomal PC/SD was also modified in comparison with non-liposomal PC experiment. The data shows an intense peak at 120 due to phase transition of amorphous viscous to liquid transparent gel and another at 225 ºC related to drug decomposition (4.5mm THK 600lb Graphite Filler Spiral Wound Gasket Ss316 API 601, curve b).Metal Hydraulic Rod Polish Buffing Equipment, through a capillary digital melting point apparatus. The curves of thermal behavior of non-Touch Screen Panel for Interactive Table, 32Open Frame IR Touch Monitor, demonstrating that SD molecules interact with phosphatidylcholine even in the dehydrated state.

CONCLUSION

Ultra-Rugged Pet Beds Orthopedic Dog Bed for Breeds Medium to Large-dried liposomes when compared to non-liposomal PC. The SD interacts with phosphatidylcholine from unilamellar liposomes modifying significantly the DSC profile, when added to either lipid or aqueous phase of liposomes. DSC analysis can be used to detect the interactions between SD and PC molecules even in dehydrated system.Convenient Use Plastic Injection Daily Use Crate Mould.

ACKNOWLEDGMENTS

The authors wish to thank FAPESP, CNPq, CAPES and PADC-FCF for financial support.

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SARBOLOUKI, M.N.; TOLIAT, T.Halal Empty Hard Gelatin Capsules Size 0 Separated and Full Avaliable(aqueous & lyophilized).PDA J. Pharm. Sci. Technol.,Bethesda, v.52, p.23-27, 1998.[Home Decoration 3D Wall Coatings Printing Form Wallpaper]

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Recebido para publicação em 03 de fevereiro de 2005
Aceito para publicação em 24 de novembro de 2006

1u 12 Port 24 Port Fiber Optic Patch PanelCorrespondence:
A.G. Oliveira
Departamento de Fármacos e Medicamentos
Faculdade de Ciências Farmacêuticas
Rodovia Araraquara-Jaú km 01
14801-902 – Araraquara-SP-10.1 Inch TFT LCD Display 1280X800 TFT LCD Matrix Module
E-mail:375W Good Quality Mini Concrete Pan Mixers