Cortical thickness across the cingulate gyrus in schizophrenia and its association to illness duration and memory performance.

Schizophrenia has been associated with structural brain abnormalities and cognitive deficits that partly change during the course of illness. In the present study, cortical thickness in five subregions of the cingulate gyrus was assessed in 44 patients with schizophrenia-spectrum disorder and 47 control persons and related to illness duration and memory capacities. In the patients group, cortical thickness was increased in the posterior part of the cingulate gyrus and related to illness duration whereas cortical thickness was decreased in anterior parts unrelated to illness duration. In contrast, cortical thickness was related to episodic and working memory performance only in the anterior but not posterior parts of the cingulate gyrus. Our finding of a posterior cingulate increase may point to either increased parietal communication that is accompanied by augmented neural plasticity or to effects of altered neurodegenerative processes in schizophrenia.

Release of gentamicin sulphate from biodegradable PLGA-implants produced by hot melt extrusion.

For a long-term local treatment of osteomyelitis biodegradable poly(lactic-co-glycolic acid) (PLGA) implants loaded with gentamicin sulphate (GS) were prepared, analysed and compared to the marketed product Septopal (Biomet, Darmstadt, Germany), which consists of polymethylmethacrylate (PMMA) beads loaded with the same active ingredient. The implants were manufactured by hot melt extrusion with a twin screw extruder. In order to decrease the processing temperature and to improve the drug release behaviour, polyethylene glycol 400 (PEG 400) was added as plasticizer in different concentrations. The glass transition temperature of PLGA measured by differential scanning calorimetry declined in the same manner as the extrusion temperature with increasing PEG 400 concentration. The extrudates of all batches exhibited good encapsulation efficiency between 85% and 115% of the specified content. The behaviour of the implants during exposure to a release medium were characterised by scanning electron microscopy, gravimetric analysis and finally in vitro drug release studies. The results suggest that drug liberation is not affected by the addition of PEG 400, and depends on the drug-PLGA ratio only. Extrudates with 25% GS showed a release pattern with an initially higher drug release followed by a zero order kinetic for about four weeks and showed release profiles equivalent to Septopal.

A new formulation concept for drugs with poor water solubility for parenteral application.

The parenteral application of active substances with poor solubility in water is often bound to the use of stabilizing excipients or surfactants with serious undesired side effects. A new concept is introduced based on a drug concentrate, comprising the active substance dissolved in parenterally acceptable organic solvents, and an aqueous dilution medium, which are mixed in a special mixing device immediately prior to application and thus generating the applicable formulation directly prior to administration. Due to the requirement of formulation stability for only a few minutes, the amount of stabilizing agents can be reduced significantly. It can be shown that model drugs dissolved in a mixture of polyoxyethylen glycol, ethanol and soya lecithin as stabilizer may be mixed to an aqueous glucose solution resulting in a parenterally acceptable and administerable dispersion which is physically stable for several minutes. First in vivo data show good tolerability and blood plasma levels which are comparable to conventional solutions.

Cyclodextrin derivatives in pharmaceutics.

The current cyclodextrin (CD) literature is reviewed concerning synthesis, characterization, and pharmaceutical relevant applications of CD derivatives. Although natural CDs have been used extensively to improve pharmaceutical properties, the effects of chemically modified CDs on the solubility, dissolution rate, and stability of drugs are overproportional. Concerning the parenteral application, the major interest is focussed on highly water-soluble, randomly substituted hydroxyalkyl derivatives of beta- and gamma-CD such as 2-hydroxypropyl-beta-cyclodextrin (2-HP-beta-CD). Although the heptakis-(2,6-di-O-methyl)-beta-cyclodextrin is applied in the pharmaceutical field, 2-HP-beta-CD is predestined as a parenteral drug carrier owing to its weak hemolytic activity and intrinsically amorphous character. A minimal average degree of substitution is especially preferred when 2-HP-beta-CD is used as solubilizer of pharmaceuticals for the use in parenteral applications. The influence of the type, degree, and pattern of substitution of the CDs, as well as substituent effects of the guest molecule is elucidated.

Piroxicam release from spray-dried biodegradable microspheres.

Poly(D,L-lactide) (DL-PLA) and poly(D,L-lactide-co-glycolide) (DL-PLGA) microspheres containing a non-steroidal anti-inflammatory model drug, piroxicam, were prepared by a spray drying process. The microspheres were characterized for surface morphology by scanning electron microscopy, particle size distribution by laser diffraction spectrometry, drug content and in vitro drug release. The diameters of the microspheres ranged from 1 to 15 microns. The DL-PLA particles appeared to be more spherical and smooth than the DL-PLGA particles, which showed a more undulated surface. Piroxicam content in the microspheres was 10%. A very high encapsulation efficiency of 99.0% was achieved with both polymers. In vitro release studies were carried out in a flow-through cell. The in vitro release rate of the drug from the DL-PLA microspheres was very slow. Less than 20% of the loaded drug was released within 10 d. The release mechanism was diffusion controlled and followed a square root of time relationship. Only a very small initial burst effect was observed. In contrast, the DL-PLGA microspheres provided a much faster drug release: about 50% was released within the first 5 h of the experiment. The mechanism for piroxicam release from the DL-PLA microspheres is not matrix erosion, but mainly drug diffusion through the intact polymer barrier. For the DL-PLGA microspheres, a pore diffusion release mechanism is proposed.

Characterization of co-polymers of lactic and glycolic acid for supercritical fluid processing.

Polymers of lactic and glycolic acid are often used for the production of injectible microparticles with controlled drug release. In the variety of processes used for the microparticle formulation, the Aerosol Solvent Extraction System (ASES) is rather special. Microparticle formation and drying take place in one step by precipitating a methylene chloride solution of the polymer in supercritical CO2. This process sets special requirements to the polymers in crystallinity, solubility, and thermal behavior that are best fulfilled by blocked copolymers. This study investigates a number of lactide-co-glycolide polymers with blocked distribution of the co-monomers by NMR spectroscopy and powder diffraction. The molar ratios are determined by 1H-NMR spectroscopy to verify the manufacturer's declarations of the purchased specimens. Additionally, the block length is determined by application of 13C-NMR. Therefore, a method reported in the literature was modified and evaluated in order to calculate the length of lactide and glycolide sequences in the polymer. Furthermore, this study looks at the impact of synthesis conditions on block length and crystallinity, and the impact of the blocking on both, crystallinity and solubility of the polymers.

Particle size, surface hydrophobicity and interaction with serum of parenteral fat emulsions and model drug carriers as parameters related to RES uptake.

Fat emulsions for parenteral nutrition, stabilized by egg lecithin, were characterized in terms of parameters relevant to uptake by the reticuloendothelial system (RES), e.g. size distribution, surface hydrophobicity and adsorption of serum components as a measure of the degree of opsonization. Adsorption of serum components was quantified by zeta potential measurement. Fat emulsions for nutrition were compared with emulsions used for drug delivery and model drug carries for intravenous injection. The emulsions for drug delivery were stabilized by the blockcopolymers Poloxamer 188 and 407 (Pluronic F68 and F127) and Poloxamine 908. Model drug carriers were hydrophobic and hydrophilic polystyrene latex particles. Hydrophilic particles were prepared by adsorption of Poloxamine 908 (coating) onto the particle surface. The hydrophobicity and serum protein adsorption decreased from hydrophobic latex particles to egg lecithin emulsions and blockcopolymer emulsions and particles. The data correlated with that in the literature concerning liver uptake in vivo showing complete RES clearance of hydrophobic latex particles, reduced uptake of egg lecithin emulsions and avoidance of RES uptake by Poloxamine 908 coated particles.

Increasing drug solubility by means of bile salt-phosphatidylcholine-based mixed micelles.

Study of the solubilization of commercial grades of soya phosphatidylcholine (SPC) with different purities by bile salts (BS) indicated that only highly pure grades of SPC are suitable for the preparation of clear solutions of BS/SPC-mixed micelles (BS/SPC-MM). The solubilizing capacity of different BS towards SPC increased in the following order; Sodium cholate (SC) < sodium deoxycholate (SDC) < sodium glycocholate (SGC). Moreover, egg phosphatidylcholine (EPC) was solubilized to a higher extent than SPC. Furthermore, the solubility study of different drugs in the prepared MM showed substantial enhancement of solubility, the extent of which is essentially affected by the chemical nature of the drug and the composition of MM. Benzodiazepine drugs such as clonazepam, tetrazepam, diazepam, and lorazepam displayed higher affinity for MM compared with BS alone, whereas steroidal drugs, such as estradiol, prednisolone and progesterone, compared with benzodiazepines, displayed relatively higher affinity for BS alone. The solubilizing capacity of MM for the different drugs was increased to different degrees by the addition of benzyl alcohol which was comparable to the solubility of the drug in pure benzyl alcohol. The interaction between benzyl alcohol and the drug in MM could be proved by NMR.

Metered-dose inhaler formulations with beclomethasone-17,21-dipropionate using the ozone friendly propellant R 134a.

Metered-dose inhalers (MDI) are the most widely prescribed devices in the treatment of lung diseases but the continued use of chlorofluorocarbons (CFC) as propellants has made them unpopular due to their influence on the stratospheric ozone layer. The purpose of this study was to show possibilities of formulating beclomethasone-17,21-dipropionate (BDP) with the alternative propellant R 134a as a solution or as a suspension-type metered-dose inhaler. Influencing factors such as surfactant concentration, cosolvent content and actuator tube design were investigated. Metered-dose inhaler formulations were manufactured using a pressure filling technique. The resulting formulations were characterized with regard to their emitted fine particle fraction using the two-stage impinger, BP 93. Fine particle fraction was found to be independent on the surfactant concentration but highly dependent on the cosolvent content and the actuator tube design. In vitro fine particle fractions of 50% were obtained with solution phase MDIs. Formulating BDP as a suspension resulted in unstable dispersions in most cases because of the partial solubility of the drug in the liquified propellant. Stable suspension formulations gave an in vitro fine particle fraction of about 30%. A comparison with established marketed BDP suspension formulations which were found to emit a fine particle fraction in the range 10-50% showed the equivalence of the new CFC-free formulations.

Size controlled production of biodegradable microparticles with supercritical gases.

Polymer microparticles were produced by means of the aerosol solvent extraction system. A solution of 3% w/w poly(L-lactic acid) in dichloromethane was sprayed into supercritical or near critical carbon dioxide gas phase. The mean particle size by volume moderately depended on the nozzle diameter and the spraying pressure used. When the polymer solution was saturated with carbon dioxide up to 5.0 MPa, the particle size and morphology of the particles were unchanged and the product was less agglomerated. Microparticles with mean diameters from 6 to 50 microns were achieved by decreasing the carbon dioxide density from 690 to 250 kg/m3. The surface structure of the particles sprayed in low density carbon dioxide showed cracks and holes. All other particles were non-porous with a smooth surface.

Mixture experiments with the oil phase of parenteral emulsions.

The effects of the oil phase as a mixture (binary, ternary) on the emulsion droplet size were investigated. The binary trials were performed with the aid of simplex lattice design with constraints. Droplet diameter was evaluated in terms of the oil phase viscosity and the interfacial tension between oil phase and the aqueous phase. As a result it could be shown that increasing the oil phase viscosity as a function of castor oil concentration led to a greater increase in particle size. At the same time, decreasing the interfacial tension of the oil phase as a function of oleic acid or oleic alcohol was shown to have a negligible effect on the particle size of the dispersed phase. A further aim was to find out a formulation by using a ternary oil phase resulting in a stable emulsion which could pass the autoclaving process. It was ascertained that oleic acid as a part of the oil phase led to proper formulation showing a satisfactory stability.

Radiolabelling of parenteral O/W emulsions by means of neutron activation.

Parenteral O/W emulsions containing lanthanide fatty acid derivatives were prepared. With regard to enhancing the incorporation efficiency of the neutron activatable excipients, the addition of the non-ionic co-emulsifier Solutol HS 15 proved to be most suitable. Comparing the different chain lengths of the fatty acids, the long chain fatty acid derivative lanthanide(tri)stearate seemed to be superior in strengthening the interfacial layer. After neutron activation, the physical and chemical stability of the irradiated formulations was evaluated. The chemical stability, indicated by the concentration of lyso phosphatidylcholine as the degradation product of the main emulsifier, was shown to be dependent on the irradiation time. By applying a neutron flux of 2.1 x 10(13) neutrons/cm2 per s, the maximum should not rise above 60 s. The physical stability indicated by the particle size distribution was affected by the presence of the non-ionic co-emulsifier. Concerning the amount of radiation necessary for in vivo biodistribution studies the maximum load of Samarium fatty acid derivatives did not yield sufficient radioactivity levels. However, Europium derivatives could be shown to be suitable for in vivo studies.

The influence of alkali fatty acids on the properties and the stability of parenteral O/W emulsions modified with solutol HS 15.

Arachis oil based parenteral O/W emulsions were prepared using soya bean phosphatidylcholine (SPC) and different combinations of co-emulsifiers containing polyethylene glycol fatty acid esters (Solutol HS 15) and alkali fatty acids (sodium laurate, sodium stearate). The parameters measured were droplet size (both by photon correlation spectroscopy and laser diffractometry), pH and zeta potential. All emulsions were subjected to autoclaving. The addition of polyethylene glycol 12-hydroxy stearate (Solutol HS 15) led to a significant decrease of mean oil droplet size. For long-term stability the amount added turned out to be the most important factor. With increased amounts of Solutol HS 15 the packing density of the emulsifier layer and the zeta potential decreased leading to instability. The optimum load of Solutol HS 15 was found to be 15 micromol/ml. Alkali fatty acids markedly improved the physical stability of the emulsions. Improved stability properties conferred to emulsions by alkali fatty acids could be attributed to the zeta potential increase even in the presence of Solutol HS 15. Consequently a mixed emulsifier film was established in which the ionized fatty acids determined the interface charge. In addition to this a strengthening of the molecular interactions occurring between phospholipid and Solutol HS 15 emulsifier in the presence of ionized fatty acids at the O/W interface can be assumed (L. Rydhag, The importance of the phase behaviour of phospholipids for emulsion stability, Fette Seifen Anstrichm. 81 (1979) 168-173). Different co-emulsifier mixtures were shown to have a pronounced impact on the plasma protein adsorption onto emulsion droplets.

Complex formation of sericoside with hydrophilic cyclodextrins: improvement of solubility and skin penetration in topical emulsion based formulations.

The main objective of this study was to devise novel methods for improving the solubility of the anti-inflammatory triterpenoid sericoside, the main component of Terminalia sericea extract, thus enabling its incorporation into topical formulations. Sericoside was stabilized by complex formation with hydrophilic derivatives of beta- and gamma-cyclodextrins in a molar ratio of 1.0:1.1. The complex of extract and cyclodextrin was equilibrated in water at 25 degrees C for approximately 24 h. The dehydrated complexes of T. sericea extract and cyclodextrin were characterized by differential scanning calorimetry, thermogravimetry analysis and X-ray diffraction. Complex formation with beta-cyclodextrin as well as gamma-cyclodextrin derivatives was detectable using these three analytical tools; however, only complexes with gamma-cyclodextrin derivatives showed stability upon storage after incorporation into topical o/w or w/o formulations. Furthermore, a T. sericea extract/gamma-cyclodextrin complex incorporated in an o/w formulation resulted in a 2.6-fold higher percutaneous penetration of sericoside in in vitro excised pig skin as compared to pure T. sericea extract. For the first time, the virtually insoluble anti-inflammatory active sericoside was incorporated into a topical emulsion based formulation in a stable manner, resulting in efficient skin penetration.

Effect of hydrotropic substances on the complexation of sparingly soluble drugs with cyclodextrin derivatives and the influence of cyclodextrin complexation on the pharmacokinetics of the drugs.

The influence of hydrotropic compounds on complex formation by 2-hydroxypropyl-beta-cyclodextrin (2-HP-beta-CD) was investigated with methyltestosterone (MeT). Various representatives of the lyotropic series were used for this purpose. Additive hydrotropic effects were observed for nicotinamide and urea, which disrupt the water structure, while structure formers such as sorbitol exerted negative effects. The effects of hydrotropic substances on the phase solubility relationship of MeT showed that inclusion complex formation with 2-HP-beta-CD depends on the degree of ordering of the solvent and is apparently subject to entropy effects. Combined systems comprising 2-HP-beta-CD and auxiliary substances with various underlying solubilizing principles were also investigated. Combination of 2-HP-beta-CD with conventional solubilizers, such as 1,2-propylene glycol or sodium deoxycholate, reduced the solubilization capacity of 2-HP-beta-CD. Competitive displacement of the inclusion molecule from its 2-HP-beta-CD complex by sodium deoxycholate suggested that cholesterol participates in the release mechanism of the inclusion molecule under in vivo conditions. The spontaneous release of complexed drug molecules could indirectly be shown on the basis of the spontaneous action of a complexed dihydropyridine derivative after iv administration in rats. The bioavailability of an investigational drug in cynomolgus monkeys could be enhanced sevenfold by inclusion complexation with 2-HP-beta-CD.

Hydroxypropyl-beta-cyclodextrin derivatives: influence of average degree of substitution on complexing ability and surface activity.

The average degree of substitution of mixtures of hydroxypropyl-beta-cyclodextrin derivatives has a large influence on the complexing abilities and physiochemical properties of the derivatives. A low degree of substitution is preferable, since these derivatives show the best complexing properties and, at the same time, low surface activities.

Complexation of steroid hormones with cyclodextrin derivatives: substituent effects of the guest molecule on solubility and stability in aqueous solution.

The inclusion complexation of homologous derivatives of steroid hormones with cyclodextrins and 2-hydroxypropyl-beta-cyclodextrin (2-HP-beta-CD) was investigated with regard to underlying structure-interaction relationship. The interaction was studied by phase solubility analysis and stabilization effects of complex formation with 2-HP-beta-CD. The solubilizing and stabilizing abilities of 2-HP-beta-CD were generally more effective for testosterone derivatives than for estradiol esters. Within a homologous series of steroid hormones, the steepest linear solubility isotherms were found for 17-methyl and 3-methyl derivatives. The solubilization of steroid esters by 2-HP-beta-CD depended on the structure and length of the ester side chain. The interaction of 2-HP-beta-CD with the steroids was hindered by long-chain fatty acid ester groups. With increasing length of the side chain, a decline of the isotherms occurred and the phase solubility behavior changed from linear to exponential. Contrary to expectations, benzoylation of steroids considerably decreased the guest-host interaction. The observed rates of degradation of the steroid esters were significantly reduced by 2-HP-beta-CD, depending on the chain length, and correlated well with the order found in phase solubility analysis. The degradation showed no deviations from pseudo-first-order kinetics, and the degradation mechanism was not changed because of complexation. The results suggest that interaction of 2-HP-beta-CD with steroid esters involves the ester functions of the prodrugs and is more suitable for unsubstituted guest molecules.

Particle size analysis of latex suspensions and microemulsions by photon correlation spectroscopy.

The particle size in microemulsions and other highly dispersed systems was determined by means of photon correlation spectroscopy (PCS). As PCS cannot be applied to highly concentrated dispersed phases, the measurement accuracy was tested for its dependence on the particle concentration using latex suspensions. The data obtained by clipping and scaling were compared. The particle size determination was expected to provide information about the influence of the structure of the surfactant system on microemulsions, using a homologous alcohol series as cosurfactant and potassium oleate as surfactant. In this system the region of solubilization is characteristically divided from the region of microemulsification by a zone of instability. Furthermore, there are distinct differences in mean particle sizes between microemulsions (9-30 nm) and micellar solutions (4-6 nm).

Particle size distributions and particle size alterations in microemulsions.

Several component diagrams for different alcohols as cosurfactants and potassium oleate as the surfactant were investigated. Between the regions of the water-in-oil microemulsion and the micellar solution (reverse micelles), and diagrams showed a zone of instability which was determined by particle size analysis by means of photon correlation spectroscopy (PCS). The polydispersity of the internal phase was determined at the instant of microemulsion formation and after fixed intervals of storage. At the edge of the microemulsion region, a rapid increase in particle size due to coalescence followed by breaking was observed. If a slow transition toward the region of solubilization could be seen by a slow decrease of the droplet size, coalescence was observed after storage. In the middle of the microemulsion region, the particle radius, however, was almost constant for a long interval. The velocity of the microemulsion formation depended on the alcohol used. In some cases a very fast formation (milliseconds) was observed; in others, macroemulsions were formed which became transparent within a few hours or days. PCS was used to follow the dynamic process of formation and breaking of such systems with droplet diameters of 5-200 nm by obtaining the mean hydrodynamic diameters. Distribution curves were calculated by the Laplace transform of the correlation function. The practicability of the method was demonstrated with mono- and polydispersed latex suspensions and microemulsions.

In vitro corneal permeability of diclofenac sodium in formulations containing cyclodextrins compared to the commercial product voltaren ophtha.

The influence of different cyclodextrin derivatives on the in vitro permeability of diclofenac sodium through pig cornea was investigated and compared to the commercial product Voltaren ophtha. (Hydroxypropyl)-beta-cyclodextrin (HP beta CD) and two amorphous methylated cyclodextrins with different degrees of substitution were used. In hemolysis studies on human erythrocytes, the hemolytic activity of the different cyclodextrins and the drug was assessed. It was shown that HP beta CD reveals the most favorable toxicological properties. A decrease in the hemolytic activity of diclofenac was yielded by adding HP beta CD. In the permeability experiments the dependency of the permeability coefficients and lag times on the type of cyclodextrin and pH of the solutions were examined. A solution containing HP beta CD buffered in the pH range 6.5 to 7 is proposed as a useful eye drop formulation. All cyclodextrin formulations showed advantages as compared to Voltaren ophtha.