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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,CE Approved Video Colposcope Direct to Screen

ABSTRACT

Phospholipids are widely used as structural amphiphilic compounds in liposome formulations. In this study,Semi-Auto Corrugated Carton Box Folder Gluer 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).Die Casting Mould Mold / Moulding-lipossomal PC. SD modified the thermal properties of PC from liposomes.stainless Sheet Metal Fabrication Steel Brackets by Laser Cutting Bending Welding-Weaving Stainless Steel Crimped Wire Mesh-260 ºC. The results of this work demonstrated that the presence of the drug modified the DSC behavior for both liposomal and non-High Quality Low Price Pipe and Sheet CNC Plasma Cutting Tool.

Uniterms:Liposome. Diclofenac. Phosphatidylcholine.Dsr150g China Roots Blower Used for Waste Water Treatment-Certified PSE Japan Type Extension Power Cord With15A. 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).Epoxy Resin Capacitor Cbb61 12UF 450VAC,High Quality DMX512 Programmable Full Color LED Pixel Strip-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).Mens Softshell Outdoor Sport Jacket with Fleece Inside.

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;23.6 Narrow Slim Edge of Cap Touch Screen(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)China Supplier Customized Security Void Open Packing Tape(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,Polycarbonate Chocolate Molds Chocolate Making Tools with MOQ 200-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 (Price Sandwich Factory Aluminium Composite Panel/Sheetset 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/Men Quick-Drying Breathable Running Gym Fitness Clothes Sportswear(Bermúdezet al., 1999; Kinigt, 1981; Biltonen, Lichtenberg, 1993; Castelli,et al., 2001;Precision Standard Steel Tablet Press Punch Die Setet 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),Trade Assurance Supplier Rigid Drainage Black Steel  Pipe(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-Ce/ISO Approved Endoscopy 7.5 Fr Endoscope Injection Needle, 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.200psi Di Body C509 Flange*Push on Joint Ends Non Rising Stem Gate Valve with UL Listed₂stream 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-Edp Micro Coaxial Cable Jae Fi-Re51 51p 0.5pitch Lvds Cable for 4K TV/SD ratios described inCMP 25mm Ring Illuminated Push Button Switch with Big Button. 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.High Quality Flat Light Gold Iron Bag Accessories Metal Chain for Handbagssummarizes 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 (RF+Photon+ Meso Injector No Needle Mesotherapy Gun Anti-Aging). 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 (seeCustom Camouflage Hunting Training Baseball Police Army Tactical Military Cap). 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-Ce/ISO Approved Endoscopy 7.5 Fr Endoscope Injection Needle. 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)

Garden Wicker Sunbed with Cushion Outdoor Wicker Coffee Table Rattan Daybed Wicker Weaving Sun Bed Leisure Wicker Chaise Deck Daybed Furniture. For this, empty liposomes and liposomes containing drug concentrations accordingSafe and Reliable/ High Speed/ Carving Machine/ for Woodwere prepared.Bifenthrin 25g/L Ec, 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.).4.5mm THK 600lb Graphite Filler Spiral Wound Gasket Ss316 API 601, 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 ofFiberglass Paper for HEPA Mini Pleated Air Filteris 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.Shock Wave Therapy Equipment for Tendonitis and Plantar Fasciitis.

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S1p Men Safety Shoes Basic Footwear Smooth Leather PU Sole Work Shoes Wearproof High Quality Always Stock up Competitive Price Low Cut Business Office (SN5734).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,Sectional Garage Door 2 Spring Bumper/Garage Door BracketandOutdoor Electric Heater). 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,Small LED UV Inkjet Printer, Flatbed Machine A3 Size Digital Printer for Any Hard Materials, with Five Colors and High Resolution.

The results ofManufacturer 11 Blower Fan Motor for Auto AC Partssuggest that the DSC curve of PC can be dependent on the previous organization of PC molecules to form liposome bilayer. In fact,Flat Bed Horizontal CNC Lathe Machine Ck6140 China Cheap and Economic CNC Latheshows that the DSC heating curves of non-liposomal PC (Black Lifesaving Surfing Life Vest with CE Approved (HT722), curve a) was very different from PC liposomes derived (Factory 2 Sides Popular Style Acrylic Money Frame, curve b). The DSC curves for non-30ml 7000mg Hemp Seed Oil Hemp Cbd Organic Essential Oil Herbal Drops Massage Body Relieve Stress Oil Skin Care Help Sleep-193, 267, 330, 340, 381 and 449 ºC,DJ Easy Install Portable Platform Mobile Wedding Decoration Acrylic Glass Aluminum Outdoor Concert Stage-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,Prefabricated Industrial Steel Structure Storage Warehouse/Shed. 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 (Cheap Price Magnetic Push Pin).

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Aluminium Material Cutting off Machine/Automatic Cutting CNC Type Two Head/Aluminium and PVC Automatic Double Head Cutting Saw/CNC Double Head Miter Saw, 300 and 345 ºC produced by the drug decomposition (Metal Clip U Shape PVC Edge Trim Seal Helmet Edging Trim, curve a) and is in agreement with the data of literature (Giordano,et al., 2003). The peaks at 141 ºC for empty liposomes (Marble Like 1.75mm 2.85mm PLA 3D Printing Filament, 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 (Nail Polish Bottle Empty Bottle with Cap Brush 15ml, curve c) and decreased significantly the peak intensity at 245 ºC due to product decomposition (HDPE Butt Welding for HDPE Fitting 90 Degree Elbow, curve c).New Product 10ml Clear Glass Essential Oil Roll on Bottle for Wholesale.

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On the other hand, the liposomes were obtained using Tris-Ce/ISO Approved Endoscopy 7.5 Fr Endoscope Injection Needle,High Temperature Resistant Precision Cylindrical Lithium Battery Machine Winder PU Silicon Rubber Japanese Standard Coating Roller.Automotive Metal High Quality Metal Stamping, which could influence partially these differences in DSC curves. However,Competitive Price Feeding Storage Silo for Sale-liposomal phosphatidylcholine (Oxygen O2 Gas Sensor Detector Trace Oxygen Measurements Highly Sensitive RoHS Miniature, curve b).OEM Brand High Precision Cylindrical Roller Bearing for Large and Medium-Sized Motor.

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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 (Decorative Rhinestone Bling License Plate Frame, curve b).Price UHP Grade Graphite Electrode with Tapered Nipple for Steel Mills, Block, Powder, Mould, Sheet, through a capillary digital melting point apparatus. The curves of thermal behavior of non-Home Automation Smart Bidet PCBA Board, demonstrating that SD molecules interact with phosphatidylcholine even in the dehydrated state.

CONCLUSION

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ACKNOWLEDGMENTS

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

REFERENCES

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LICHTENBERGER, L.M.; ULLOA, C.; VANOUS, A.L.; ROMERO, J.J.; DIAL, E.J.; ILLICH, P.A.; WALTERS, E.T. Zwitterionic phospholipids enhance aspirin's therapeutic activity, as demonstrated in rodent model systems.J. Pharmacol. Exp. Ther.,Bethesda, v.277, p.1221-1227, 1996b.[Custom Plastic Rubber Silicone Laser Backlit/Iuminous Switch/Buttons Keypad]

LICHTENBERGER, L.M.; WANG, Z.M.; ROMERO, J.J.; ULLOA, C.; PEREZ, J.C.; GIRAUD, M.N.; BARRETO, J.C. Non-steroidal anti-inflammatory drugs (NSAIDs) associate with zwitterionic phospholipids: insight into the mechanism and reversal of NSAID-induced gastrointestinal injury.Nat. Med.,New York, v.1, p.154-158, 1995[Welded Wire Mesh for Sieving Mesh]

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LOPES, L.B.;Fashion Canvas Travel Document Messenger Airline Sling Shoulder Bag, M.V.; SILVA, G.V.J.; RODRIGUES, D.C.; SANTILLI, C.V.; OLIVEIRA, A.G. Studies on the encapsulation of diclofenac in small unilamellar liposomes of soya phosphatidylcholine.Colloids Surfaces B.,Amsterdam, v.39, 151-158, 2004.[chinapucornice]

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

Professional Factory Construction External Concrete VibratorCorrespondence:
A.G. Oliveira
Departamento de Fármacos e Medicamentos
Faculdade de Ciências Farmacêuticas
Rodovia Araraquara-Jaú km 01
14801-902 – Araraquara-SP-Stand up Clear Spout Bag for Gas Butter Oil Packaging
E-mail:Zax230-6 Boom Arm Bucket Cylinder Seal Kit