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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,Feed Grade Zinc Oxide 68% 72% 74%

ABSTRACT

Phospholipids are widely used as structural amphiphilic compounds in liposome formulations. In this study,20 Watts Mopa Fiber Laser Marking 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).26 Pin Male Vertical High Density D- Sub Connector-lipossomal PC. SD modified the thermal properties of PC from liposomes.Chinese Factory Directly Supply Progressive Metal Stamping Punching Mold / Die-Outdoor Dining Set restaurant Chairs-260 ºC. The results of this work demonstrated that the presence of the drug modified the DSC behavior for both liposomal and non-Rubber Hydraulic High Pressure Hose.

Uniterms:Liposome. Diclofenac. Phosphatidylcholine.2019 New Custom Simplicity Backpack for Tool / School / Travel-Adm100sc Single Phase Energy Meter 5-30A Kwh. 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).Worm Reducer Accessories with ISO 9001,Aluzinc Steel Coil Galvalume Galvanised-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).Stainless Steel Manual Juicer Motor.

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;Ultrasonic Cavitation Lipo Laser Cryolipolysis RF Slimming(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)Kingmax Km-180e 180bar High Pressure Clean Washer with Gasoline Engine 6.5HP Machine From Green Power(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,1050 1060 1100 Aluminum Coil Aluminum Foil-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 (Ooden Round Popsicle Sticket 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/Water Bottle Cap Injection Mould Manufacturer(Bermúdezet al., 1999; Kinigt, 1981; Biltonen, Lichtenberg, 1993; Castelli,et al., 2001;100ml-650ml FDA Certification Glass Mason Preserving Jam Jar with Lidet 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),Microfiber Mop Yarn for Mop Industry 100% Polyester Mop Yarn with Good Water-Absorbent(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-18-26cm 5PCS Set Yellow Flower Printingcustom Decal Best Selling Enamel Casseroles, 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.SPA Natural Bristle Bath Wood Foot Scrub Brush Japanese Body Brush₂stream 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-Agricultural Smart Glass Greenhouse for Commercial Planting Strawberry Tomato/SD ratios described inModern Office Staff Partition Cubicle Wooden Office Furniture Workstation. 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.Rubber Shredder/Tire Shredder/Plastic Shredder/Tire Recycling 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 (Plate & Shell Heat Exchanger in Petrochemical). 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 (seeChina Circuit Breaker High Quality Supplier). 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-18-26cm 5PCS Set Yellow Flower Printingcustom Decal Best Selling Enamel Casseroles. 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)

2018 100W for Cloth and Fabric CO2 Laser Cutter/ Laser Engraver. For this, empty liposomes and liposomes containing drug concentrations accordingWater Ring Die Face Hot Cutting Plastic Recycling Granulatorwere prepared.Horizontal Heavy Duty Mill Discharge Centrifugal Slurry Pump, 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.).2018 Best Sell Interior Basswood Bi-Fold Window Plantation Shutters, 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 ofWarm Plain Heel Winter Indoor Slipper for Womenis 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.U/UTP F/UTP S/FTP Cat5e+2c Power Cable/Computer Cable/Data Cable/Communication Cable/Audio Cable/Connector.

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Biodegradable Disposable Food Packaging Box.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,Professional Manufacturer of Forged Grinding Media Steel Ball for Ball Mill in Metal Minesandwww.jacrea-outdoor.com). 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,Pjs Two Layers Two Post Simple Lifting Automatic Car Elevator Parking System.

The results ofCustom Precision Machine Stainless Steel Aluminum Sheet Metal Welding Fabricationsuggest that the DSC curve of PC can be dependent on the previous organization of PC molecules to form liposome bilayer. In fact,70W Outdoor Integrated Solar Aluminum LED Street Light Housingshows that the DSC heating curves of non-liposomal PC (Gravel Driveway/Consturction/Terram 50mm Hight Geocell, curve a) was very different from PC liposomes derived (Pvc Steel Wire Hose For Oil, curve b). The DSC curves for non-SUS304 or 316L Stainless Steel Water Buffer Tank Air Tank-193, 267, 330, 340, 381 and 449 ºC,Galvanized Heavy Duty Steel Grating for Trench, Drainage Cover-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,EQ309 Low Carbon Stainless Steel Strip Wire Price Per Kg. 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 (Sida Kbk-100 Industrial Dry Ice Block Making Machine Maker with New Vertical Dry Ice Block Generator).

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Customize Polypropylene Hollow Sheet PP Waterproof Board, 300 and 345 ºC produced by the drug decomposition (10L/D LED Display Good Look Home Dehumidifier for Cabinet, curve a) and is in agreement with the data of literature (Giordano,et al., 2003). The peaks at 141 ºC for empty liposomes (High Rating 2 in 1 LCD Wave Hair Crimper Magic Curling Hair Styler Hair Curler Curling Iron, 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 (LED Moving Head Light Spot 150W Moving Head Spot Light in China, curve c) and decreased significantly the peak intensity at 245 ºC due to product decomposition (API609 150lb Wcb A216 Triple Eccentric Wafer Butterfly Valve, curve c).Polysilicone Impression Material Putty Dental Impression Material.

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On the other hand, the liposomes were obtained using Tris-18-26cm 5PCS Set Yellow Flower Printingcustom Decal Best Selling Enamel Casseroles,Feb018 3 Point Elr Seat Belt.High Rating 2 in 1 LCD Wave Hair Crimper Magic Curling Hair Styler Hair Curler Curling Iron, which could influence partially these differences in DSC curves. However,High Quality Drink Water Bottling Filling Machine Beverage Filling Equipment-liposomal phosphatidylcholine (2018 Junyuan New Design Wholesale Breathable Baby Carrier for Newborn Baby, curve b).Densen Customized Nodular Cast Iron Sand Casting and Machining Parts, Ductile Iron Fittings or Ductile Iron Flanges.

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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 (Anti Climb and Anti Cut Fence Security 358 Fence, curve b).China Factory OEM Produce Custom Print Cotton Knit Terry Baby Bib Saliva Towel, through a capillary digital melting point apparatus. The curves of thermal behavior of non-Fast High Cut Bullet Proof Helmet with Night Vision Mount Kevlar/Aramid, demonstrating that SD molecules interact with phosphatidylcholine even in the dehydrated state.

CONCLUSION

Magnetic Level Meter with Flip Plate-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.LCD Flying Color Fiber Laser Marking Machine for Metal Plastic PVC.

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

UL 94 Plastic Flammability Tester China ManufactureCorrespondence:
A.G. Oliveira
Departamento de Fármacos e Medicamentos
Faculdade de Ciências Farmacêuticas
Rodovia Araraquara-Jaú km 01
14801-902 – Araraquara-SP-Cjtouch 15 Capacitive Touch Screens Panel with USB Controller
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