Hydrogel matrices containing single and mixed natural cyclodextrins. Mechanisms of drug release

Hydrogel networks of α, β or γ-cyclodextrin (CD) and mixtures of α/β or β/γ CDs have been obtained using epichlorohydrin (EP) as a crosslinking agent. Discs of the resulting polymers were evaluated as drug carriers for controlled release using the antiinflammatory naproxen (NAP) as a model drug. βCD polymer (βCDP) has shown the highest amount of drug loaded and the lowest one corresponds to the polymer containing αCD, in agreement with the affinities of NAP for the corresponding cyclodextrins.     

Characterization and release behaviors of porous PCL/Eudragit RS microcapsules containing tulobuterol

In this work, porous poly(ɛ-caprolactone) (PCL)/Eudragit RS 100 (ERS-100) microcapsules containing tulobuterol base as a model drug were prepared by a solvent evaporation method and the effect of the quaternary ammonium groups of ERS-100 on the release behaviors of the microcapsules was investigated. The microcapsules prepared with PCL alone showed a stable and smooth surface, whereas porous microcapsules were formed with the addition of ERS-100. Drug loading and encapsulation efficiency of the microcapsules were slightly decreased with an increase of ERS-100 content, resulting from an increase in the porosity of the microcapsules. In an acidic release medium, PCL microcapsules showed slow drug release, whereas PCL/ERS-100 microcapsules showed a faster release rate with an increasing ERS-100 content. These behaviors are likely due to an increase in the diffusion rate of the drugs stemming from an increased hydration of the microcapsules, which results from the interaction between the carboxyl group of the release medium and the quaternary ammonium group of ERS-100.     

Retention properties of cyclodextrins and modified cyclodextrins in reversed phase HPLC

Summary In the course of systematic studies on the solubility, hydrophobicity and complexation properties of cyclodextrins and modified cyclodextrins, the retention behavior of , , and of some glycosylated cyclodextrins has been examined by means of reverse phase HPLC. Mobile phase mixtures containing large amounts of water have been used because of the possible application of such systems to biological studies. Mobile phase mixtures with both methanol and acetonitrile show a linear relationship between the volume fraction of the organic part of the mobile phase and the logarithm of the capacity factor. The extrapolation of capacity factors to a total aquous system are used and compared to other techniques (including solubility) in order to evaluate the hydrophobic properties of the cyclodextrins. In particular, the solubility of cyclodextrins has been explored for a wide range of organic solvent/water mixtures. Whilst cyclodextrins are definitely the most hydrophobic, followed by glycosylated cyclodextrins, the others behave differently in the two mobile phase systems. The differences observed in the results are related to the chemical nature of the organic phase. Comparisons between chromatographic and solubility methods are given and interpretations are proposed. Some cyclodextrins have been modified to increase or modify not only the hydrophobicity but also the solubility, the complexation and the molecular recognition of drugs. The most important aim of this study was to define conditions and rules for further drug vectorization by cyclodextrin-drug complexation.     

Study of aerodynamic and release properties of inhaled particles containing cyclodextrins

The aim of this study was to investigate the influence of different cyclodextrins (CyD) on physical characteristics of inhalation dry powders. The particle size was characterised by Aerosizer^® LD and aerodynamic behaviour of inhaled complexes assessed by twin-stage liquid impinger. The in vitro release profile of the powders was studied through Franz cell modified method. Produced particles showed a suitable size for pulmonary delivery, ranging between 1 and 5 μm. The nature of the CyD affected the powders performance on reaching the lower compartment (“Lungs”), mainly by the altering their aerodynamic properties, which is reflected on the different percentages of their emitted respirable fractions. HP-γ-CyD:fluticasone propionate complex showed a fast release of corticosteroid while γ-CyD had a constant release throughout time. The best characteristics for pulmonary delivery were obtained with acetyl-γ-CyD:fluticasone propionate complex.     

Effect of preparation temperature in solvent evaporation process on Eudragit RS microsphere properties

Eudragit RS 100 microspheres containing ketoprofen as a model drug were prepared by the solvent evaporation method using an acetone/liquid paraffin solvent system. The influence of various preparation temperatures: 10, 25, 35, and 40 degrees C, on particle size and morphology, drug content and release kinetics, and drug crystal state was evaluated. With increasing temperature, microsphere average size was found to increase and particle size distribution to widen significantly. At 10 degrees C particles of irregular shape are formed, whereas higher temperatures gradually improve the sphericity of microspheres. As can be seen from SEM photographs, particle surface roughness decreases as preparation temperature increases. It was found that temperature had no effect either on ketoprofen microencapsulation efficiency or on its crystal state, but it does influence emulsion-stabilizer incorporation. Ketoprofen forms solid solution in Eudragit matrix and maintains amorphous state for significant period of time. Drug release rates from microspheres correlated with microspheres' surface roughness and to a lesser extent with particle size.     

Release properties of chemical and enzymatic crosslinked gelatin-gum Arabic microparticles containing a fluorescent probe plus vetiver essential oil

Oil-containing gelatin-gum Arabic microparticles were prepared by complex coacervation followed by crosslinking with glutaraldehyde or transglutaminase. A fluorescent mixture, khusimyl dansylate (KD) as the fluorescent compound mixed to the vetiver essential oil, was used as oil model. The effect of the type of crosslinking of the coacervated gelatin-gum Arabic membrane, the physical state of microparticles, wet or freeze-dried and the type of release media, aqueous with surfactants, Sodium Dodecyl Sulphate (sds) or Tween 80 (tw) and anhydrous ethanol as organic media on the release rate of the KD from the microparticles, was experimentally investigated.It was shown that the oil was dispersed uniformly throughout the microparticles and the chemical crosslinked microparticles were more resistant to swelling, presenting smaller sizes after hydration. Also the crosslinking effect, transglutaminase or glutaraldehyde, could be confirmed by the integrity of the crosslinked gelatin-gum Arabic microparticles after incubation in the aqueous sds media, compared to complete dissolution of the uncrosslinked microparticles in this media.The cumulative fluorescent KD release from the gelatin-gum Arabic microparticles decreased in the following order of dissolution media: anhydrous ethanol > tw > sds and the wet microparticles have shown a faster KD release than freeze-dried ones. A mathematical model was used to estimate the diffusion coefficient (D). The chemically crosslinked gelatin-gum Arabic microparticles ensured a pronounced retard effect in the KD diffusion, presenting a D varying from 0.02 to 0.6 × 10⁻¹¹ cm²/s, mainly in an aqueous media, against D varying from 1.05 to 13.9 × 10⁻¹¹ cm²/s from the enzymatic crosslinked microparticles.     

Preparation and characterization of inclusion complexes containing fixolide, a synthetic musk fragrance and cyclodextrins

AHTN (7-Acetyl-1,1,3,4,4,6-hexamethyl-1,2,3,4-tetrahydronaphthalene), commercially known as fixolide or tonalide, is a synthetic fragrance widely used in replace of natural musk odor which is more expensive. It is a popular fragrance material added in the manufacturing of personal care and household products, such as perfumes, soaps, shampoos, detergents, and fabric softeners. AHTN is semivolatile and is degraded under light exposure and high temperature. This work focuses on the complexation of AHTN with cyclodextrins in the effort to stabilize the fragrance material. AHTN was complexed with β-cyclodextrin, methyl (MβCD), and hydroxypropyl (HPβCD) derivatives in the mole ratio 1:1, 1:2, and 1:3 guest:host, and the complexes formed by physical mixing, co-precipitation, kneading, and freeze-drying were analyzed by DSC and FTIR. Percent AHTN included in the complex was also determined by hexane extraction and GC analysis. It was found that no inclusion complex was formed in the physical mixture. When co-precipitation method was performed, only βCD could form inclusion complex with AHTN, while the other two derivatives could not. Using 1:2 AHTN:βCD, no free AHTN was left in the complex as evidenced by DSC and FTIR spectrum. In kneading and freeze-drying methods, complexes could be formed with all CDs tested. However, co-precipitation method with 1:2 AHTN:βCD and kneading method with 1:2 AHTN:MβCD provided the highest complex yield with highest amount of AHTN included in the complex. AHTN in the complex form was more stable against high temperature and UV exposure than its free form.     

Effect of polysaccharide nanocrystals on structure, properties, and drug release kinetics of alginate-based microspheres

Polysaccharide nanocrystals, such as rod-like cellulose nanocrystals and chitin whiskers and platelet-like starch nanocrystals, were incorporated into alginate-based nanocomposite microspheres with the aim of enhancing mechanical strength and regulating drug release behavior. The structures and properties of the sols and the resultant nanocomposite microspheres were characterized by rheological testing, Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD) and scanning electron microscopy (SEM). The presence of polysaccharide nanocrystals increased the stability of the crosslinked network structure, and the nanocomposite microspheres consequently exhibited prominent sustained release profiles, as demonstrated by inhibited diffusion of theophylline. Furthermore, based on the drug release results, the release kinetics and transport mechanisms were analyzed and discussed.     

Separation of peptides and proteins by capillary electrophoresis using acidic buffers containing tetraalkylammonium cations and cyclodextrins

A method for improving separations of peptides and other positively charged species in capillary zone electrophoresis with untreated capillaries using acidic buffers containing tetraalkylammonium cations is described. Tetramethylammonium and tetrabutylammonium cations dynamically modify the capillary surface, leading to a reversal in the direction of the electroosmotic flow. As a result, the adsorption of positively charged peptides and proteins is minimized, and resolution and peak capacity are improved as the migration of cationic analytes is counterbalanced by the electroosmotic flow. The combining effect of reversing electroosmotic flow and cyclodextrin inclusion complexation on separations of closely related peptides and a protein mixture, as well as tryptic digest of hemoglobin is demonstrated.     

Effect of secondary polymer on self‐precipitation of pH‐sensitive polymethylmethacrylate derivatives Eudragit E100 and Eudragit L100

Interpolyelectrolyte complexes or polyplexes can be seen as interesting alternatives in the purpose of active ingredients encapsulation. Working on polymethylmethacrylate derivatives with special focus on controlled oral drug delivery, the influence of charged polyelectrolytes (polyacrylic acid, polyethylenimine, and amino‐dextran) and noncharged ones (polyvinyl alcohol, dextran 40, and Pluronic F68) has been investigated on the precipitation of two pH‐sensitive Eudragit polymers, namely, L100 and E100. Moreover, the possibility of preparing polyplexes involving the two polymethylmethacrylate derivatives with different charged and noncharged secondary polyelectrolytes has been studied. The obtained dispersions have been characterized in terms of mean particle size, size distribution, zeta potential, and morphology. Direct precipitation of Eudragit L100 by medium acidification in a batch process and in the presence of polyethylenimine allowed the production of particles with a narrow size distribution. The mean size was around 200 nm. In this case, the zeta potential was found to be +45 mV at pH = 7 in 1mM aqueous NaCl solution, and the produced suspension was stable in time since no aggregation and sedimentation have been observed. A precipitation pH of 8.16 allows us to suggest the preparation of a polyplex based on Eudragit L100 and polyethylenimine. In contrary, polyvinyl alcohol has shown ability to induce an increase in particle mean size whereas other polyelectrolytes showed no significant effect. Moreover, it was observed that polyethylenimine and polyacrylic acid solutions were able to directly induce Eudragit E100 precipitation whereas amino‐dextran and noncharged polyelectrolytes showed no effect on its precipitation and on particle size distribution.     

Effect of oxygen plasma treatment on the release behaviors of poly(epsilon-caprolactone) microcapsules containing tocopherol

The biodegradable poly(epsilon-caprolactone) microcapsules (PCL) containing tocopherol (TC) were prepared by emulsion solvent evaporation method, and microcapsules were treated by oxygen plasma to enhance the hydrophilic microcapsules. The morphologies and thermal properties of the microcapsules were determined by SEM and DSC measurements. The microcapsules studied were characterized by surface free energy or work of adhesion through contact angle measurement. As a result, the features of the microcapsules could be adjusted by manufacturing condition, such as surfactant and core ratio. The surface free energy or work of adhesion of the microcapsules was increased with increasing the time of plasma treatment, which could be attributed to the increased hydrophilic groups during oxygen plasma treatment. The release profile of the microcapsules was determined by UV-vis spectroscopy and the microcapsules containing tocopherol showed the rapid release rate, as compared with untreated ones.     

Amphipathic and membrane-destabilizing properties of the cationic acrylate polymer Eudragit E100

The cationic acrylate polymer Eudragit E100 (E100) produces a biphasic effect on the stability of casein micelles disrupting their internal structure. These results suggested that this polymer could have some amphipathic character. Therefore, in this study the polymer was characterized with respect to its interaction with different amphipathic systems (bile-acid micelles, lipoproteins and liposomes), cell membranes (red blood cells) and virus membranes (Herpes simplex type 2 virus). As with caseins, a biphasic effect was observed with bile acids with a precipitation phase at low polymer/bile acid ratio and a solubilization phase when the polymer concentration was increased. Upon interaction with human plasma, an important reduction in cholesterol and triglycerides was observed upon remotion of E100 by a rise in pH to 8.5 and centrifugation. In agreement with this finding, an important reduction in plasma lipoproteins was observed upon its treatment with E100 and further remotion by pH rise and centrifugation. However, the amount of the major protein components of human plasma and the activity of several enzymes and antibodies were not affected by their treatment with E100. The membrane-destabilizing properties of E100 were confirmed by its lytic activity on liposomes and red blood cells and by an important antiviral effect of E100 on Herpes simplex virus type 2. Altogether, these results show that, despite its water solubility and cationic character, E100 displays a significative amphipathic and membrane-destabilizing character with potential biotechnological applications. [diagram in text].     

Influence of substituent position and cavity size of the regioisomers of monocarboxymethyl‐α‐, β‐, and γ‐cyclodextrins on the apparent stability constants of their complexes with both enantiomers of Tröger's base

Enantiomers of Tröger's base were separated by capillary electrophoresis using 2^I‐O‐, 3^I‐O‐, and 6^I‐O‐carboxymethyl‐α‐, β‐, and γ‐cyclodextrin and native α‐, β‐, and γ‐cyclodextrin as chiral additives at 0–12 mmol/L for β‐cyclodextrin and its derivatives and 0–50 mmol/L for α‐ and γ‐cyclodextrins and their derivatives in a background electrolyte composed of sodium phosphate buffer at 20 mmol/L concentration and pH 2.5. Apparent stability constants of all cyclodextrin–Tröger's base complexes were calculated based on capillary electrophoresis data. The obtained results showed that the position of the carboxymethyl group as well as the cavity size of the individual cyclodextrin significantly influences the apparent stability constants of cyclodextrin–Tröger's base complexes.     

Liposomal drugs dispersed in hydrogels. Effect of liposome, drug and gel properties on drug release kinetics

Release of calcein and griseofulvin (GRF) from control (gels in which solutes are dissolved in) and liposomal gels was studied using agarose-assisted immobilization as a technique to separate gels from drug-receptor compartments. Liposomes composed of phosphatidylcholine (PC) or distearoyl-glycero-PC and cholesterol (DSPC/Chol), and incorporating calcein or GRF were prepared by thin film hydration. After cleaning the liposomes they were dispersed in different hydrogels (carbopol 974 [1, 1.5 or 2% (w/w)], hydroxylethyl-cellulose (HEC) [4% (w/w)], or a mixture of the two), and release of calcein or GRF was followed by fluorescence or photometric technique, respectively. Results show that calcein release from liposomal gels is slower compared to control gels, and can be further retarded by using rigid-membrane liposomes (faster release from PC-liposome compared to DSPC/Chol-liposome gels). Additionally, calcein release is not affected by the lipid amount loaded (in the range from 2 to 8 mg/ml), therefore solute loading can be controlled according to needs.

Oppositely, GRF release from liposomal gels is determined by drug loading. At high drug loading levels (compared to GRF aqueous solubility), GRF is released with constant rate from liposomal gels irrespective of liposome type (PC or DSPC/Chol). Thereby, for amphiphilic/lipophilic drugs, drug properties (solubility, log P) determine the system behavior.

Calcein and GRF release from control carbopol gels is faster compared to HEC and mixture gels. The same is true for calcein in liposomal gels. Carbopol gel rheological properties were found to be significantly different (compared to the other gels), implying that these characteristics are important for drug diffusion from gels.     

Effect of the reaction temperature on the transglycosylation reactions catalyzed by the cyclodextrin glucanotransferase from Bacillus macerans for the synthesis of large-ring cyclodextrins

The synthesis of cyclodextrins with from 6 to more than 50 glucose units by cyclodextrin glucanotransferase (CGTase, EC 2.4.1.19) from Bacillus macerans was investigated. Analysis of the synthesized cyclic α-1,4-glucan products showed that a higher yield of large-ring cyclodextrins were obtained with a reaction temperature of 60 °C compared to 40 °C. The yield of large-ring cyclodextrins obtained at 60 °C represented about 50% of the total glucans employed in the reaction. Analysis of the cyclodextrin-forming cyclization reaction and of the coupling reaction of the CGTase resulting in the degradation of mainly the larger cyclic α-1,4-glucans indicated higher rates of the cyclization reaction at 60 °C compared to 40 °C while the opposite was found for the coupling reaction.     

Study of the interaction between cyclodextrins and liposome membranes: effect on the permeability of liposomes

We wanted to compare and understand the effect of the most currently used cyclodextrins on a model membrane. We studied the influence of most currently used cyclodextrins on the release of a fluorescent marker encapsulated in the inner cavity of SUV liposomes. It was shown that the observed effect on calcein release can be directly related to the affinity of cyclodextrins for both lipid components of liposomes, cholesterol and phosphatidylcholine. From this relationship, we were able to determine, for each cyclodextrin, a theoretical concentration giving rise to 50% or 100% calcein release. This theoretical concentration was confirmed experimentally.     

Purification of glutathione reductase from chicken liver and investigation of kinetic properties

Glutathione reductase was purified from chicken liver and some characteristics of the enzyme were investigated. The purification procedure was composed of four steps: preparation of homogenate, ammonium sulfate precipitation, 2′,5′-ADP Sepharose 4B affinity chromatography, and Sephadex G-200 gel filtration chromatography. Owing to the four consecutive procedures, the enzyme was purified 1714-fold, with a yield of 38%. Specific activity at the final step was 120 enzyme unit (EU)/mg of protein. The purified enzyme showed a single band on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weight of the enzyme was found to be 100 kDa by Sephadex G-200 gel filtration chromatography, and the subunit molecular weight was found to be 43 kDa by SDS-PAGE. Optimum pH, stable pH, optimum ionic strength, and optimum temperature were 7.0, 7.4, 0.75 M Tris-HCl buffer including 1 mM EDTA, and 50°C, respectively. K _M and V _max values for NADPH and glutathione disulfide (GSSG) substrates were also determined for the enzyme.     

Study on the inclusion behavior and solid inclusion complex of 5-amino-6-methyl-2-benzimidazolone with cyclodextrins

The inclusion behaviors of three native or modified CDs including p-CD,2-hydroxypropyl-β-CD(2-Hp-β-CD) and 2,6-dimethyl-β-CD(Me-β-CD) toward 5-amino-6-methyl-2-benzimidazolone(AMBI) were comparatively investigated by NMR and fluorescence titration in combination with IR spectra,X-ray diffractometry and scanning electron microphotographs.The experimental results jointly demonstrated that the phenyl ring of AMBI entered into the cavity of the CDs and located close to the narrow rims accompanied by the formation of the 1:1 inclusion complex with large stability constant in aqueous solution.The introduction of the hydroxypropyl unit to the host improved the solubility,ultimately effecting an obvious promoting in the fluorescence intensity and the stability constant     

Effect of ethanol on the synthesis of large-ring cyclodextrins by cyclodextrin glucanotransferases

Cyclodextrin glucanotransferase (EC 2.4.1.19, CGTase) synthesizes cyclodextrins (CD) composed of 6 to more than hundred glucose units from amylose by an intramolecular transglycosylation reaction. The addition of ethanol to the reaction medium resulted in an increase of the yield of large-ring CD obtained with a CGTase from Bacillus sp. BT3-2 and Bacillus macerans. The presence of 15% ethanol in the reaction mixture with the CGTase from Bacillus sp. BT3-2 resulted in a 30% increase of the amounts of CD₁₀–CD₁₃ synthesized after 5 h of reaction. The addition of 20% ethanol increased the yield of CD₁₄–CD₂₁ up to 1000%. The hydrolysis of the large-ring CD by the CGTases was suppressed in the presence of ethanol. The ring-opening coupling cyclization reactions of the CGTase were effected differently by the organic solvent which may contribute to the observed increase of the yield and size of the CD obtained in the synthesis reactions.     

Influence of substrate and product concentrations on the production of cyclodextrins by CGTase of Bacillus firmus,strain no. 37

The influence of substrate or product level on the initial velocity of cyclodextrin (CD) production by cyclodextringlycosyltransferase from a Brazilian isolate of Bacillus firmus was studied. Our results indicate that the product γ-CD is a stronger inhibitor to the reaction than β-CD. Small saccharides could also inhibit CD production, although to a lesser extent than the products, and maltose was the strongest inhibitor among small saccharides. Increasing substrate concentration resulted in greater reduction on enzyme activity for the formation of β-CD than for γ-CD. We modeled the kinetics of CD production with a set of four reversible reactions including the cyclization/coupling reaction that forms/opens CDs, and three disproportionation reactions. Our model on the initial velocity data explained well the substrate inhibition phenomenon. Kinetic parameters were determined by fitting the initial velocity data into our model.