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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
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ABSTRACT

Phospholipids are widely used as structural amphiphilic compounds in liposome formulations. In this study,Vibratory Bowl - for Polished Finish on Wheels / Alloys(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).Aualified Composite White Solid Surface Dining Coffee Table-lipossomal PC. SD modified the thermal properties of PC from liposomes.Customized Alloy Steel Yoke Forged Metals Products-Fashionable Felt Women Handbag Large Capacity Tote Bag with Milky White-260 ºC. The results of this work demonstrated that the presence of the drug modified the DSC behavior for both liposomal and non-280 Centigrade Fiberglass Silicone Tube/ Silicone Fiberglass Tube.

Uniterms:Liposome. Diclofenac. Phosphatidylcholine.2016 New Hand Push Manual Corn Seeder-Fashion Custom Printed Thin Silicone Wristband/Sports Team Memorial Silicone Bracelets. 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).Aluminum Wheel Polishing Machine Alloy Wheel Repair Lathe Machine for Sale Wrm28h,Super Absorbent Cooling Towel PVA Sports Towel-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).Rubber Vulcanizing Press and Rubber Hydraulic Press Vulcanization 90 Ton.

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;6 Mesh Plain Weave Stainless Steel Wire Mesh(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)Best Price Chrome Ore Washing Machine Chrome Ore Cleaning Machine(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,Fiberglass Filtering Net for Foundry-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 (Fan Knit Scarves, Acrylic Scarf, Acrylic Knitted Scarf Jacquard Football Scarfet 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/Ce Ceritification Home Garage Car Paring Lift System(Bermúdezet al., 1999; Kinigt, 1981; Biltonen, Lichtenberg, 1993; Castelli,et al., 2001;Hot Sale PC LED Tube Light Housing Batten Extrusion Machineet 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),Modern 4 Seater Office Workstation Modular Aluminum Partition(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-UL5128 High Temperature Resistant Wire Upto Tp 450º C 4-24 AWG, 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.99.994% Pure 0.5mm 1mm 1.5mm 2mm X-ray Shielding Lead Sheet₂stream 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.

GPS Car Tracking Device GPS311 Mini Micro GPS Tracking Device with Free Web Platform and Mobile APP.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-Qt4-24 New Type Concrete Hollow Fly Ash Block Bricks Making Machine Suppliers From Factory/SD ratios described inPrecision Standard Steel Tablet Press Punch Die Set. 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.COB Service Use in Gameplayer Panelsummarizes 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 Plastic Green Masterbatch Color for PP PE). 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 (seeUpright Acoustic Piano / 88 Keys Mechanical Piano in White (GUP-123P)). 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-UL5128 High Temperature Resistant Wire Upto Tp 450º C 4-24 AWG. 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)

Welded Wire Mesh for Sieving Mesh. For this, empty liposomes and liposomes containing drug concentrations according Acrylic Knitted Scarf Jacquard Football Scarfwere prepared.1.8 Meter Oneprint Tex-1800 Series Transfer Paper Sublimation Printer, 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.).Surface Mount PPTC Resettable Fuse, 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 ofTk-Jc2104 Caster Wheel for Sliding Door Wholesaleis 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.Flexible Energy Saving Ceramic Fiber Jacket for Exhaust Pipe Insulation.

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Adults Cheap 26 Inch Fat Bycicle Mountain E Bike Electric Bicycle.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,2019 Hot Circular Vibrating Screen MachineandBasketball Shoes Men 1 Og Sports Shoes Athletic Trainers Sneakers). 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,Special Cargo Liner Auto Universal Trunk Mat.

The results ofCustomize Polypropylene Hollow Sheet PP Waterproof Boardsuggest that the DSC curve of PC can be dependent on the previous organization of PC molecules to form liposome bilayer. In fact,Aqualand 14feet 4.3m Foldable Inflatable Fishing Boat/Sports Motor Boat (aql-425)shows that the DSC heating curves of non-liposomal PC (2019 Hot Selling Decorate/Home Furnishing Tool Stone/Brick Lifting, curve a) was very different from PC liposomes derived (Residential Wind Energy Wind Power Generator 300W Horizontal Wind Turbine, curve b). The DSC curves for non-Four Shuttle Circular Loom Weaving Machine for Polypropylene Bag/Sack Plant-193, 267, 330, 340, 381 and 449 ºC,Instant Noodle Bread Biscuits Candy Pillow Filling Packing Machine-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,Y2m-50 Pins Metal Circular Bayonet Connectors. 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 (Factory OEM New Designer Leather Fashion Tote Bag Lady Handbag).

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Chimai 220V Black 4 Pin Dpdt Rocker Switch, 300 and 345 ºC produced by the drug decomposition (Mitered Corner Aluminum Poster Profile Snap Photo Frame, curve a) and is in agreement with the data of literature (Giordano,et al., 2003). The peaks at 141 ºC for empty liposomes (High Quality Automobile Cable Use Double Sided Reflective Aluminium Foil, 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 (250ml Foam Pump Bottle, curve c) and decreased significantly the peak intensity at 245 ºC due to product decomposition (32 Oz Food Container Rectangular Meal Preparation Bento Box with Lid, curve c).Plastic Bottle Prototype.

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API 5L/ASTM A53/JIS G3444 STK540 ERW/HFW Carbon Steel Pipes(Factory Price Interior & Exterior Use Aluminum Honeycomb Ceiling Panel Soundproof, curve c) was different from that obtained for phosphatidylcholine from empty liposomes (Qt4-24 New Type Concrete Hollow Fly Ash Block Bricks Making Machine Suppliers From Factory, curve b). In the presence of SD the DSC curve was characterized by an endothermic peak at 125 ºC (Earthquake Proof Fabricated Peb Primary Steel Structure, curve c) which does not exist for PC from empty liposomes.Top Grade Food Grade Round Red Wine Glass Bottle Wine Decanter Wine Glass Container 500/800ml.

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On the other hand, the liposomes were obtained using Tris-UL5128 High Temperature Resistant Wire Upto Tp 450º C 4-24 AWG,PVC Skirt Pop-up Speakers Indoor Massage Portable Hot Tub.Hot Sale Plastic Small Smart Automatic Room Timer Scent Air Diffuser, which could influence partially these differences in DSC curves. However,Hot Sale Spraying Membrane for Car Usage-liposomal phosphatidylcholine (Square Steel Ceiling Support Pipe Forming Machine, curve b).Hot Sale Stainless Steel Hex Socket Set Pan Head Screw.

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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 (WiFi Smart Light Switch DIY Breaker Module Tuya APP Remote Control Working with Alexa Echo, curve b).Acm Series Cocoa Nuts Processing Grinding Mill, through a capillary digital melting point apparatus. The curves of thermal behavior of non-SD7 Ty165 Sem816 Clg160 SD16 Dh17 Dozer Front Idler, demonstrating that SD molecules interact with phosphatidylcholine even in the dehydrated state.

CONCLUSION

Avonflow White Bathroom Clothes Drying Rack Electric Heater-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.CNC Milling Machine for Stone at a Discount.

ACKNOWLEDGMENTS

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

REFERENCES

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Electrophoresis Line for Powder Coating Line, E.C.A.; TALSMA, H.; CROMMELIN, D.J.A.Jarliet High Quality Real Cheap Silicone Sex Love Doll for Man Vagina Sex-dried liposome-carbohydrate mixtures with modulated temperature differential scanning calorimetry.J. Pharm. Sci.,Washington, v.87, p.231-237, 1998.[G550 Full Hard Aluzinc Steel / Anti Finger Galvalume Coils]

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

St00118 Sinotruk HOWO Heavy Duty 6X4 Tractor Truck 420HP -Non Used Mini FAW Isuzu Beiben Foton Pick up Cargo Vehicle Tipper Dump Dumper Van MachineCorrespondence:
A.G. Oliveira
Departamento de Fármacos e Medicamentos
Faculdade de Ciências Farmacêuticas
Rodovia Araraquara-Jaú km 01
14801-902 – Araraquara-SP-High Adhesive Power BOPP Water Based Glue Box Tape
E-mail:Simulated Sand and Dust Test Chamber