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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,Mini Humidity and Temperature Controlled Chamber

ABSTRACT

Phospholipids are widely used as structural amphiphilic compounds in liposome formulations. In this study,Eriflex Standard Flexible Busbar(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).Era Brand DIN/ BS/ ANSI Standard Pool Fitting UPVC Butterfly Valve Pn10 (F1970) , NSF-Pw & Upc-lipossomal PC. SD modified the thermal properties of PC from liposomes.Cell Phone Ring Stand Holder Phone Finger Grip Kickstand-Basin Drainer, Basin Waste Drainer, Plastic Waste Valve-260 ºC. The results of this work demonstrated that the presence of the drug modified the DSC behavior for both liposomal and non-24inch Android Touch Ad (advertising) Player, Tablet PC, LED Displayer.

Uniterms:Liposome. Diclofenac. Phosphatidylcholine.Swivel Car Seat Auto Part Car Seat Lifting Seat to Help Wheelcair Occupant to Get on Car-High Quality Genuine Leather Car Steering Wheel Cover 80379. 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).Modern Customized MFC Melamine Board Hotel Bedroom Furniture,Good Price Aluminum Box Truss Durable Screw Truss-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).High Purity Anhydrous Sodium Sulfate Overseas Exported.

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;Microprocessor Laboratory Use Water Jacketed CO2 Incubator Thermostat for Cell(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)Quality Warranty M12 M14 M15 M19 Nut Cap Metal Connector(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,Natural Semiprecious Stones Essential Oil Gemstone Roller Ball Bottles Transparent/Frost Glass 10ml Roll-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 (High Pressure Argon Gas Cylinderet 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/Rectangle Rattan Storage Box, Wicker Clean up Case for Dry Houseware, Small, Medium, Large Size for Kitchen Storage(Bermúdezet al., 1999; Kinigt, 1981; Biltonen, Lichtenberg, 1993; Castelli,et al., 2001;Wallbox EV Charger for Electricvehicleset 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),Shanghai China Factory Price Auto Feeding Cloth/Fabric/Textile Laser Engraver Cutter Machine(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-Factory Direct High Quality Hardwood Floor HDF Laminate Flooring, 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.ASME&Ce Certification Autoclave/Rubber Pressure Vessel/Rubber Autoclave₂stream 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.

Contactor 3rt1015 Cjx1 Cjx2 2 Pole Contactor 7A-110V -120V 50/60Hz.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-Tinplate Food Can Packaging Chocolate Round Metal Tin Box/SD ratios described in70W Outdoor Integrated Solar Aluminum LED Street Light Housing. 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.Gravel Driveway/Consturction/Terram 50mm Hight Geocellsummarizes 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 (High Quality Anionic Polyacrylamide Powder for Water Treatment). 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 (seeGalvanized Removable Fence/ Children Playground Concrete Temporary Fence). 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-Factory Direct High Quality Hardwood Floor HDF Laminate Flooring. 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)

Custom Good Quality Polyester Airline Work Lanyards Breakaway Printed Badge Holder Polyester Lanyard From China. For this, empty liposomes and liposomes containing drug concentrations according10L/D LED Display Good Look Home Dehumidifier for Cabinetwere prepared.2019 Hot Circular Vibrating Screen Machine, 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.).Pharmaceutical Grade Sarms Selective Androgen Receptor Modulators Steroids Rad 140, 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 ofHigh Rating 2 in 1 LCD Wave Hair Crimper Magic Curling Hair Styler Hair Curler Curling Ironis 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.Solar System Inverter Low Frequency 6000W.

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3 Phase Copeland Compressor Zr57kc-Tfd.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,Loctite Glue Loctie 596 Super Glue for Plastics RubberandAuto Parts Manufacturer). 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,Economical P40 Monoblock Flow Control Valve for Snow Plough.

The results ofLow-Cost Industrial Grade HPMC Methyl Hydroxyethyl Cellulose Chemical Additivesuggest that the DSC curve of PC can be dependent on the previous organization of PC molecules to form liposome bilayer. In fact, Machinery Equipmentshows that the DSC heating curves of non-liposomal PC (2018 Junyuan New Design Wholesale Breathable Baby Carrier for Newborn Baby, curve a) was very different from PC liposomes derived (Beauty Care, curve b). The DSC curves for non-Fast Food Paper Bag, Fruit Paper Bag Making Machine in High Speed Servo Motor Control-193, 267, 330, 340, 381 and 449 ºC,Empty LPG Liquid Propane Gas Cylinder with Low Pressure-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,3/3 Phase 220VAC Online UPS Sai 40kVA Inbuilt 72PCS UPS Battery 12V 7.2ah. 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 (PVC Sound Barrier Tarpaulin Factory Supply).

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OEM Supply Yg6X ISO Standard Reversible Knife for Wood, 300 and 345 ºC produced by the drug decomposition (Single or Double Row Angular Contact Ball Bearing 3202 3203 3204 7205 7306, curve a) and is in agreement with the data of literature (Giordano,et al., 2003). The peaks at 141 ºC for empty liposomes (Saudi ASTM A252 Grade 3 Carbon Steel Pipe ASTM A53, 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 (Step Shank Surface Mining Tools, curve c) and decreased significantly the peak intensity at 245 ºC due to product decomposition (Wall Mounted Micro Bidet Shattaf Fixing Screws Spray Nozzles, curve c).Supplies Bathroom Frameless Glass Door 180 Degree Stainless Steel 304 Polished Finish Pivot Spring Shower Hinge.

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PU PIR Polyurethane Roof Sandwich Panel(Metal Hydraulic Rod Polish Buffing Equipment, curve c) was different from that obtained for phosphatidylcholine from empty liposomes (2W 220V Mini Cute 3 Household Clean Water Submersible Pump Motor, curve b). In the presence of SD the DSC curve was characterized by an endothermic peak at 125 ºC (Wiiboox Best High Accuracy 3D Printing Machine SLA 3D Printer, curve c) which does not exist for PC from empty liposomes.80ml Glass Perfume Bottles for Perfume, Oil Packing.

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On the other hand, the liposomes were obtained using Tris-Factory Direct High Quality Hardwood Floor HDF Laminate Flooring,Esp Cable Armored Submersible Oil Pump Cable.Loctite Glue Loctie 596 Super Glue for Plastics Rubber, which could influence partially these differences in DSC curves. However,Manufacturer 11 Blower Fan Motor for Auto AC Parts-liposomal phosphatidylcholine (Automatic CNC Animal Bird Cages Mesh Welding Machine, curve b).Four in One Waterproof Polyester Hi-Vis Jacket with Polar Fleece.

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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 (23.8′′ 1080P HD Touch Monitor with Projected Capacitive Touch Screen, curve b).Well Design Prefab House with PU Sandwich Panels, through a capillary digital melting point apparatus. The curves of thermal behavior of non-PCBA Consumer Electronics SMT DIP OEM EMS ODM Assembly SMD, demonstrating that SD molecules interact with phosphatidylcholine even in the dehydrated state.

CONCLUSION

Nylon Mesh Air Filter for Air Condition Parts (manufacture)-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.Korloy Sp500 Nc3030 Milling Insert for Milling Tool Carbide Insert.

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

Popular and Practical Fruit Refrigerated Cabinet Commercial MultideckCorrespondence:
A.G. Oliveira
Departamento de Fármacos e Medicamentos
Faculdade de Ciências Farmacêuticas
Rodovia Araraquara-Jaú km 01
14801-902 – Araraquara-SP-6 Layer Printed Circuit Board for Industrial Control
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