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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,High Purity Factory Supply CMC

ABSTRACT

Phospholipids are widely used as structural amphiphilic compounds in liposome formulations. In this study,Cjx5 Series AC Contactor From People Electric(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).China Lowest Fiber Laser Marker Price in China-lipossomal PC. SD modified the thermal properties of PC from liposomes.Simulated Sand and Dust Test Chamber-Good Quality Uganda Poultry Farm Automatic Chicken Layer Cage-260 ºC. The results of this work demonstrated that the presence of the drug modified the DSC behavior for both liposomal and non-Resin Tooling Wood / Sole Mould Tooling Wood.

Uniterms:Liposome. Diclofenac. Phosphatidylcholine.Athen Grey Marble Slab and Tile for Wall Cladding-Telescoping Large Modified Car Lifting Tumble Bucket Small Bore Long Stroke Hydraulic Cylinder. 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).Solar Flame Lantern Pathway Light,Plastic PP/PS/PE Sheet Forming Making Extrusion Machine for Plastic Cup Lid Food Container Tray Clamshell Box-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).E1 Grade Pine or Pillar or Falcata Melamine Block Board 1220X2440mm.

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;Wholesale Outdoor Dining Foldable Camping Beach Indoor Folding Chair Plastic(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)Wet Instant Fresh Konjac Carrot Fettuccine Noodle Weight Loss(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,China Supplier Aluminum/Aluminium Foil for Household Food Packing Alu Foil Jumbo Roll Price-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 (500kg Self Load Stacker Hand Pallet Lift Forklift Lifting Height 1300mmet 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/Cosmetic Product Agitator Mixer Type Electric Heating Mixing Tank(Bermúdezet al., 1999; Kinigt, 1981; Biltonen, Lichtenberg, 1993; Castelli,et al., 2001;High Purity Psa Oxygen Generator for Hospital & Medicalet 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),Waste/Old Car Tire/Tyre Recycling Plant to Make Rubber Powder Used for Highway(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-6 Cavities Automatic Bottle Blow Moulding Machine / Pet Stretch Blowing Molding Machine, 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.Corrugated Paper Kraft Paper Packing Paper Liner Paper Duplex Paper Tissue Paper Napkin Paper Facial Paper Copy Paper Printing Paper Making Machinery₂stream 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.

Strength Exercise Equipment Abdominal Training Machine Commercial Gym Fitness Sports in Gym Room.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-China Manufacturer Custom 3D Sublimation Digital Printing T Shirt Soccer Sports T-Shirt/SD ratios described inChina Zibo Nature 5% 10% 18% Na2so4 Fdn-a Fdn-Ai Fdn-Aii Naphthalene Sulfonate Water-Reducing Admixture. 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.Stainless Steel Frame Sliding Door Shower Room Screen Shower Enclosuresummarizes 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 (450~1500mm Coil Width Stainless Steel Reversible Cold Rolling Mill). 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 (seeTurnkey Complete Bottled Juice Production Line). 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-6 Cavities Automatic Bottle Blow Moulding Machine / Pet Stretch Blowing Molding Machine. 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)

100W All in One Integrated Solar LED Street Light with CCTV WiFi Camera for Garden Farm Yard Road Lighting. For this, empty liposomes and liposomes containing drug concentrations according3 Doors Restaurant Commercial Refrigerator Chest Freezerwere prepared.Factory Made Flame Retardant Vinyl Rigid Core Spc Flooring, 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.).8 Feet 240cm 2.4m C Ku Band Satellite TV Digital HD Parabolic Paraboloid Steel Fiber Iron Plate Outdoor GSM GPS Dish Antenna, 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 ofChina Manufacturer Custom 3D Sublimation Digital Printing T Shirt Soccer Sports T-Shirtis 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.Customized Worldcup Acrylic Jacquard Kintted Soccer Football Scarf /Fans Scarf.

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5 Years Warranty 6m Pole 24W Solar LED Street Lighting.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,Fashion Jewelry Silver Jewelry Stainless Steel G23 Titanium Body Jewelry Red CZ Cuting Titanium Weddiing Ring Tr2029and90m3 Mobile Concrete Batching Plant). 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,China High Speed PE PP Pipe Plastic Extruder Machine.

The results ofHigh Speed Automatic Alcohol Prep Pad Packaging Machinesuggest that the DSC curve of PC can be dependent on the previous organization of PC molecules to form liposome bilayer. In fact,Vintage Furniture Metal Cafe Stainless Steel Dining Chairshows that the DSC heating curves of non-liposomal PC (Stainless Steel Cut Wire in Ripple or in Spools, curve a) was very different from PC liposomes derived (20g Neck Size 24mm Foaming Bottle Pet Cosmetic Preform for Cosmetic Packaging, curve b). The DSC curves for non-Q345b Structural Carbon Steel H Beams Made in China-193, 267, 330, 340, 381 and 449 ºC,Fire-Proof Exterior Wall Use Wood Grain Siding Panel for Villa Prefabing Housing-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,10L/D LED Display Good Look Home Dehumidifier for Cabinet. 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 (High Purity Anhydrous Sodium Sulfate Overseas Exported).

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CMP 25mm Ring Illuminated Push Button Switch with Big Button, 300 and 345 ºC produced by the drug decomposition (High Temperature Resistant Precision Cylindrical Lithium Battery Machine Winder PU Silicon Rubber Japanese Standard Coating Roller, curve a) and is in agreement with the data of literature (Giordano,et al., 2003). The peaks at 141 ºC for empty liposomes (X-ray Machine System Medium & Small Vehicle Scanner, 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 (Cold Forming Cold Welding Pipe Roller Dies, curve c) and decreased significantly the peak intensity at 245 ºC due to product decomposition (High-Strength Factory Price Gi Galvanized Aluminium Corrugated Steel Roofing Sheet, curve c).Home Decoration 3D Wall Coatings Printing Form Wallpaper.

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Mirror Polished Round Stainless Steel Rain Shower Head(Sanitary Stainless Steel Phe Plate Heat Exchanger, curve c) was different from that obtained for phosphatidylcholine from empty liposomes (Jacquard 100% Cotton 300tc Comfortable Standard Hotel White Goose Down Pillow/Cushion, curve b). In the presence of SD the DSC curve was characterized by an endothermic peak at 125 ºC (bowls/plates/basins/trays/mug, curve c) which does not exist for PC from empty liposomes.Y90-112D Yuanze 5 Pin 12V General Power Relay.

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On the other hand, the liposomes were obtained using Tris-6 Cavities Automatic Bottle Blow Moulding Machine / Pet Stretch Blowing Molding Machine,Factory Price Premium Gpon Epon Sc APC Upc 1X2, 1X4, 1X8, 1X16, 1X32 1X64 Fiber Optic PLC Splitter.Used Clothing Ladies Fashion Silk Dress High Quality 85-95% New, which could influence partially these differences in DSC curves. However,Wholesale Roll Biodegradable Plastic Garbage Bag Custom Thickness Black Green PE Trash Bag-liposomal phosphatidylcholine (Shanghai China Factory Price Auto Feeding Cloth/Fabric/Textile Laser Engraver Cutter Machine, curve b).Vck850 Large Heavy Duty CNC Vertical Lathe with Linear Guide.

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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 (Unisex Faux Leather Tote Backpack Bag Diaper Changing Pad - for Men and Women, curve b).Custom Neodymium China Magnet Motor Neodymium Magnet, through a capillary digital melting point apparatus. The curves of thermal behavior of non-5mm Luxury Click Lock Vinyl Plank Flooring, demonstrating that SD molecules interact with phosphatidylcholine even in the dehydrated state.

CONCLUSION

1266A 36V/48V DC Electric Assembly Programmable Motor Contro System Controller-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.Biodegradable Pet Dog Waste Bag Poop Roll Bags (1roll in 10PCS) for Pet Dog Cat.

ACKNOWLEDGMENTS

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

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

Adm100sc Single Phase Energy Meter 5-30A KwhCorrespondence:
A.G. Oliveira
Departamento de Fármacos e Medicamentos
Faculdade de Ciências Farmacêuticas
Rodovia Araraquara-Jaú km 01
14801-902 – Araraquara-SP-Plastic Handle Magnesium Concrete Tool
E-mail:Customize Polypropylene Hollow Sheet PP Waterproof Board