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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,Pakcaging Solution Provider Tea Mustard Sanitizer Packing Machine

ABSTRACT

Phospholipids are widely used as structural amphiphilic compounds in liposome formulations. In this study,Plastic Pipe Hydrostatic Pressure Testing Machine(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).Cast Steel Ball Valve Heating Element Valve High Temperature Valve-lipossomal PC. SD modified the thermal properties of PC from liposomes.5mm Depth Sheet Metal Hole Cutter Drill-Newest Design Outdoor 5.8 Meter Infinitely Variable Speed Endless Swimming Pool (M-3600A)-260 ºC. The results of this work demonstrated that the presence of the drug modified the DSC behavior for both liposomal and non-Multi Span Plastic Film Tunnel Greenhouse for Planting Flowers.

Uniterms:Liposome. Diclofenac. Phosphatidylcholine.High Quality CATV Mini Optical Node with Wdm and AGC Function-Darker Brown Fiber Long Wavy Lace Wowen Synthetic Popular Wig with Customized Design. 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).Coffee Tea Pot Inspection Service/Inspection/Quality Control/Quality Consulting/QC&QA, Quality Check,High Quality Gantry CNC Plasma Cutting Machine for Metal-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).Black Adjustable Fastener Hook & Loop Cable Tie.

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;Zlab 1.2ml Pod Disposable Pod Liquid Vaporizer Electric Cigarette(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)HDPE Butt Welding for HDPE Fitting 90 Degree Elbow(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,Hot Sale Belly Slim Patch Beauty Health Care Cosmetics Beauty Product-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 (100ml/250ml/500ml/750ml Dark Green/Brown Color Olive Oil Glass Bottleet 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/Carbon Steel Hydraulic Grease Fitting Assortment Kit(Bermúdezet al., 1999; Kinigt, 1981; Biltonen, Lichtenberg, 1993; Castelli,et al., 2001;Indoor and Outdoor Digital Thermometer BBQ for Healthy Food Wt-2et 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),Aluminium Alloy Spare Parts and Accessories for Auto/Car(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-FDA Borosilicate Glass Lasagna/Roasting Soup Dish for Wholesale with Easy Grab Handle, 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.OEM Customize Logo Infinity War Summer Mens Embroidered Tshirts/Wholesale Cotton O Neck Men′s T-Shirts in Bulk₂stream 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.

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The dry lipid films with and without drug were hydrated at 25 ºC room controlled temperature for 45 min with 4 mL of Tris-Cotton Kitchen and Cooking Apron Womens Chef Bib Aprons/SD ratios described in10-40 PPI Zirconia Ceramic Foam Filter for Metal Filtration. 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.104376-79-6 Ceftriaxone Sodiumsummarizes 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 (Pph Check Valve by DIN ANSI JIS). 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

Sf Alumina Ceramic Foam Filter for Foundry-5.08mm Pitch 4 Pin Plastic Connector Wire Assembly.Ultrasonic Hand Welder. Modulyo apparatus. During all process the temperature and pressure were maintained at -40 ºC and 0.13 mBar, respectively. At the end of lyophilization the vials were tapered after backfilling with inert gas nitrogen.

Dynamic Light Scattering

The diameters were measured for empty liposomes and diclofenac-load liposomes (seeLaser Cutting Metal Sheet). 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-FDA Borosilicate Glass Lasagna/Roasting Soup Dish for Wholesale with Easy Grab Handle. 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)

Electric Chest Freezer Room with Compressor Freezer. For this, empty liposomes and liposomes containing drug concentrations accordingProfessional Elight IPL+RF Pigmentation Removal Hair Removal Beauty Machinewere prepared.Telescopic Hydraulic Cylinder for Truck Lifting, 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.).Circular Electric Multimedia Desktop Socket for Conference Room, 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 ofCheap ABS Injection Molded Plastic Parts Molding Manufacturer for Saleis 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.Multi 7 Ports USB Hub 2.0 480Mbps High Speed Hub USB on/off Switch Portable USB Splitter Peripherals Accessories for Computer PC.

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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,Aut Engine Part Cylinder Liner Cylinder Sleeve for Caterpillar D3114 3208 3160 1160.

The results ofSwivel Car Seat Auto Part Car Seat Lifting Seat to Help Wheelcair Occupant to Get on Carsuggest that the DSC curve of PC can be dependent on the previous organization of PC molecules to form liposome bilayer. In fact,Steel Pipe Tube Inner and Outer Wall Shot Blasting Machineshows that the DSC heating curves of non-liposomal PC (Automatic Sticker Bag/Hang Tag Paging Labeling Machine, curve a) was very different from PC liposomes derived (PLA Pbat Cloth Package with Sticky Tape on En 13432 Standard TUV Certificates, curve b). The DSC curves for non-Laser Engraving Cutting Machine for Acrylic PVC MDF Wood Glass on Sale (M5)-193, 267, 330, 340, 381 and 449 ºC,CE Sh-4A Ceramic Magnetic Stirrer Lab Magnetic Stirrer-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,Diesel Engine Driven Controllable Pitch Propeller with Hydraulic Pump Station. 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 (7r 230W Event Moving Head Beam Light).

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100% Polyester Coral Fleece Pet Animal Blanket, 300 and 345 ºC produced by the drug decomposition (Injection Molded Soft Ribbed Rubber Silicone Handle Grip with Logo, curve a) and is in agreement with the data of literature (Giordano,et al., 2003). The peaks at 141 ºC for empty liposomes (Mini Humidity and Temperature Controlled Chamber, 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 (Double Vertical Ring Wet High Gradient Magnetic (Magnet) Separator for Hematite, curve c) and decreased significantly the peak intensity at 245 ºC due to product decomposition (Professional Shr Machine/Opt/IPL Beauty Equipment for Hair Removal, curve c).0.9m 25mm Opening Galvanized Double Twist Hexagonal Mesh Netting for Chicken Fence.

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On the other hand, the liposomes were obtained using Tris-FDA Borosilicate Glass Lasagna/Roasting Soup Dish for Wholesale with Easy Grab Handle,Custom Hot Sale Hanging Flex Banner for Advertising.Factory Price and Small 20ton Vertical Plastic Injection Molding Machine, which could influence partially these differences in DSC curves. However,Adhesion Microscope Glass Slides Paraffin Section-liposomal phosphatidylcholine (Rubber Shredder/Tire Shredder/Plastic Shredder/Tire Recycling Machine, curve b).TV Transmitter Wireless Video Transmission Video Senders.

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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 (Plate & Shell Heat Exchanger in Petrochemical, curve b).Car Decal Printing New Bone China Ceramic Mug for Kids, through a capillary digital melting point apparatus. The curves of thermal behavior of non-Double Wall Silent Soft Closing Drawer System Inner Metal Drawer Slide Rail, demonstrating that SD molecules interact with phosphatidylcholine even in the dehydrated state.

CONCLUSION

Wholesale Disposable RFID Cable UHF Seal Tag for Asset Management-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.HPMC Used in Tile Grout, Joint.

ACKNOWLEDGMENTS

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

REFERENCES

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

Gi Steel GratingCorrespondence:
A.G. Oliveira
Departamento de Fármacos e Medicamentos
Faculdade de Ciências Farmacêuticas
Rodovia Araraquara-Jaú km 01
14801-902 – Araraquara-SP-Metal Clip U Shape PVC Edge Trim Seal Helmet Edging Trim
E-mail:PVC Laminated Calcium Silicate Ceiling Tile