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In vitro characterization of jet-milled and in-situ-micronized fluticasone-17-propionate.

Particle properties are decisive for therapeutic efficiency of an inhaled pulmonary drug. Jet-milling as the common way for micronization of inhaled powder drugs shows several disadvantages such as a non-homogeneous particle size distribution and unnatural, thermodynamically-activated particle surfaces causing a high agglomeration behavior. For pulmonary use in a dry powder inhaler (DPI) beside a small particle size, a good de-agglomeration activity is required. In this study, fluticasone-17-propionate (FP) is in-situ prepared in a respirable particle size by a controlled crystallization technique. First, the drug is dissolved in acetone and precipitated by a solvent change method in the presence of a cellulose ether (HPMC) as stabilizing hydrocolloid. By rapidly pouring the drug solution into the polymer-rich water phase, the previously molecularly dispersed drug is associated to small particles and stabilized against crystal growth simultaneously by the presence of the hydrophilic polymer. This dispersion was then spray-dried. The mean particle size of the drug was around 2 microm and consequently in the respirable range. The physico-chemical properties of the in-situ-micronized drug were compared to those of an unmilled and a jet-milled quality. Differences in the X-ray patterns and amorphous parts could be detected for the jet-milled but not for the in-situ-micronized drug. In addition, the aerodynamic behavior of the engineered and the jet-milled FP was analyzed using the FlowCaps inhaler as delivery device and compared to the commercial product Flutide Diskus. The fine particle fraction (FPF) (<5 microm) was increased four-fold from approximately 9% for the jet-milled drug to approximately 40% for the in-situ-micronized drug when the pure drug powder was dispersed with the FlowCaps device.

Androstadienes↗

Gamma irradiation for terminal sterilization of 17beta-estradiol loaded poly-(D,L-lactide-co-glycolide) microparticles.

17beta-Estradiol-loaded microparticles using poly-(D, L-lactide-co-glycolide) polymer (PLG) were prepared by a modified spray-drying method and the effects of gamma-irradiation on drug substance, polymer and microparticles were investigated. Irradiation doses ranging from 5.1 to 26.6 kGy were applied using a 60Co-radiation source. 17beta-Estradiol drug substance showed excellent stability against gamma-irradiation in the investigated dose range, whereas microencapsulated estradiol seems to be converted to conjugation products with PLG, and to a lesser extent to the degradation product 9,11-dehydroestradiol. The weight-average molecular weight of the PLG polymers decreased with increasing irradiation dose while polydispersity indices (M(w)/M(n)) remained nearly unchanged, compatible with a random chain scission mechanism in lactide/glycolide-copolymer degradation. In vitro drug release studies showed accelerated kinetics with increasing irradiation doses due to dose dependent polymer degradation. Microbiological process monitoring showed decreasing bioburden with increasing spraying time, which was successfully further reduced by applying irradiation sterilization. Microencapsulated test spore suspensions of Bacillus pumilus ATCC 27142, the official test specimen for the gamma-sterilization process, revealed effective reduction of bioburden, confirming its published D(10) value. In conclusion, our studies demonstrated efficacy of gamma-irradiation as terminal sterilization method for poly-(D,L-lactide-co-glycolide) polymer-based drug delivery systems. The sterilization conditions need to be carefully adjusted for the final dosage form.

Bacillus↗

Formulation design of double-layer in the outer shell of dry-coated tablet to modulate lag time and time-controlled dissolution function: studies on micronized ethylcellulose for dosage form design (VII).

The dry-coated tablet with optimal lag time was designed to simulate the dosing time of drug administration according to the physiological needs. Different compositions of ethylcellulose (EC) powder with a coarse particle (167.5 microm) and several fine particles (< 6 microm), respectively, were mixed to formulate the whole layer of the outer shell of dry-coated tablets. The formulations containing different weight ratios of coarse/fine particles of EC powders or 167.5 microm EC powder/excipient in the upper layer of the outer shell to influence the release behavior of sodium diclofenac from dry-coated tablet were also explored. The results indicate that sodium diclofenac released from all the dry-coated tablets exhibited an initial lag period, followed by a stage of rapid drug release. When the mixture of the coarse/fine particles of EC powders was incorporated into the whole layer, the lag time was almost the same. The outer shell broke into 2 halves to make a rapid drug release after the lag time, which belonged to the time-controlled disruption of release mechanism. When the lower layer in the outer shell was composed of 167.5 microm EC powder and the upper layer was formulated by mixing different weight ratios of 167.5 microm and 6 microm of EC powders, the drug release also exhibited a time-controlled disruption behavior. Its lag time might be freely modulated, depending on the amount of 6 microm EC powder added. Once different excipients were respectively incorporated into the upper layer of the outer shell, different release mechanisms were observed as follows: time-controlled explosion for Explotab, disruption for Avicel and spray-dried lactose, erosion for dibasic calcium phosphate anhydrate, and sigmoidal profile for hydroxypropyl methylcellulose.

Cellulose↗

Behavior of enterococci in egg processing operations.

Since enterococci were detected in dried and frozen egg products (whole egg and egg yolk), the origin of the enterococci and their behavior in the different stages of egg processing were surveyed. The bacterial survey of unwashed and washed eggs, gathered from several parts of Japan, showed the presence of an average of 60 enterococci per egg on the shell surface of unwashed eggs. Smaller numbers of enterococci were detected on the shell surface of washed eggs. Most of the detected enterococci were Streptococcus faecalis and S. faecalis var. liquefaciens. The contents of two eggs were contaminated with enterococci when 120 washed eggs were examined. Enterococci were destroyed to some extent by the disinfectants used for washing the shell eggs, but they seemed to have greater resistance than Escherichia coli and Pseudomonas fluorescens used for comparison. In whole egg and egg yolk, enterococci grew rapidly at 25 C, slowly at 10 C, and very slowly at 2.5 C. The frozen storage of both whole egg and egg yolk at -20 C for 3 months only slightly decreased the number of enterococci. They were decreased only slightly by normal pasteurization but were destroyed in the desugaring process by the glucose oxidase method. Enterococci numbers were only slightly decreased by spray-drying whole egg and yolk. Considering these characteristics of enterococci, it appears to be difficult to produce enterococci-free egg products in a normal production line.

Animals↗

Pharmacokinetics and tissue distribution of ipriflavone, an isoflavone derivative, after intravenous administration to rabbits.

Pharmacokinetic parameters of ipriflavone and its main metabolites, M1 and M5, after intravenous administration of spray-dried ipriflavone, SIP (10, 20, and 30 mg/kg as ipriflavone) and tissue distribution of ipriflavone, M1, and M5 after intravenous administration of SIP (20 mg/kg as ipriflavone) were evaluated in rabbits. Saturable metabolism of ipriflavone were observed after intravenous administration; at an ipriflavone dose of 30 mg/kg, the dose-normalized (based on 10 mg/kg) AUC was significantly greater (72.4 and 64.0 versus 103 microg min/mL), Cl was significantly slower (138 and 156 versus 97.6 mL/min/kg), and terminal half-life (94.8 and 129 versus 211 min) and mean residence time (91.3 and 116 versus 186 min) were significantly longer than those at 10 and 20 mg/kg. The AUC of M1 was also significantly greater at ipriflavone dose of 30 mg/kg. The terminal half-life, AUC, and renal clearance of M5 were also significantly different at ipriflavone dose of 30 mg/kg than those at 10 and 20 mg/kg. Ipriflavone was widely distributed in most rabbit tissues studied and the tissue-to-plasma (T/P) ratios of ipriflavone were greater than unity in all tissues (or organs) studied except spleen, indicating that ipriflavone has high affinity to rabbit tissues studied, and this could be supported by considerably high values of the apparent volume of distribution of ipriflavone at steady state (11 400-16 900 mL/kg). M1 and M5 were also detected in most rabbit tissues with considerable amount of M1 (T/P ratio of 9.43) and M5 (T/P ratio of 4.66) in the kidney.

Analgesics↗

Design and validation of an annular shear cell for pharmaceutical powder testing.

An annular shear cell was constructed for powder flow testing, the influence of design and process parameters was characterized, and the results were compared with other flow methods. The shear cell was designed with interchangeable parts to mimic other shear cells. The texture of the powder-metal interface and the gap distance between the lid and side wall of the trough were varied, and the effects of shear rate, powder bed thickness, and consolidation times were tested. Shear parameters, such as cohesion, angles of friction, and flow factors, were measured for microcrystalline cellulose, anhydrous lactose, spray-dried lactose, mannitol, dibasic calcium phosphate dihydrate, anhydrous theophylline, and theophylline monohydrate powder. The results were then compared with the Carr index, mass flow rate, and flowability index. Design parameters such as surface texture and the gap distance significantly affected the shear call results, whereas for the process parameters studied, the shear rate, consolidation time, and powder bed height had a minimal effect on the shear cell results. Of the shear parameters obtained, the angles of friction best represented the known flow properties of powders and were in general agreement with those from other flow tests.

Cellulose↗

Hydrogels formed by crosslinked poly(vinyl alcohol) as sustained drug delivery systems.

Poly(vinyl alcohol) was crosslinked with ethylene glycol diglycidyl ether to obtain hydrogel-forming polymers. The polymers were also substituted with oleoyl chloride, providing hydrogels with weak solubility. These new polymeric materials were evaluated for the formulation of sustained drug delivery systems. Vancomycin hydrochloride was used as a peptidic model drug whose sustained release should minimize its inactivation in the upper part of the gastrointestinal tract. Spray-dried mixtures of the drug and the polymer [at 1:4 and 1:8 (w:w) ratios] were prepared and the release of the drugs from the mixtures was evaluated in vitro at pH 2.0, 5.5, 7.4, and 8.0. The results indicated that the crosslinked polymers slowed down the release of the drugs with respect to the pure drug at each pH. The degree of crosslinking of ethylene glycol diglycidyl ether and the extent of substitution with oleoyl chloride were found to influence drug release.

Anti-Bacterial Agents↗

Hydrogels Formed by Crosslinked Poly(vinyl alcohol) as Sustained Drug Delivery Systems.

Poly(vinyl alcohol) was crosslinked with ethylene glycol diglycidyl ether to obtain hydrogel-forming polymers. The polymers were also substituted with oleoyl chloride, providing hydrogels with weak solubility.These new polymeric materials were evaluated for the formulation of sustained drug delivery systems. Vancomycin hydrochloride was used as a peptidic model drug whose sustained release should minimize its inactivation in the upper part of the gastrointestinal tract. Spray-dried mixtures of the drug and the polymer [at 1:4 and 1:8 (w:w) ratios] were prepared and the release of the drugs from the mixtures was evaluated in vitro at pH 2.0, 5.5, 7.4, and 8.0. The results indicated that the crosslinked polymers slowed down the release of the drugs with respect to the pure drug at each pH.The degree of crosslinking of ethylene glycol diglycidyl ether and the extent of substitution with oleoyl chloride were found to influence drug release.

Journal Article↗

Cyanide utilization and degradation by microorganisms.

Various microorganisms can produce (cyanogenesis) or degrade cyanide. They degrade cyanide either to detoxify it, or to use it as a source of nitrogen for growth. Significant amounts of cyanide are formed as a secondary metabolite by a wide range of fungi and a few bacteria by decarboxylation of glycine. When cyanide has been formed by the snow mould fungus it is degraded by conversion to carbon dioxide and ammonia via an unknown pathway. In contrast, cyanogenic bacteria either do not further catabolize cyanide or they convert it into beta-cyanoalanine by addition to cysteine or O-acetylserine. Several non-cyanogenic fungi that are pathogens of cyanogenic plants are known to degrade cyanide by hydration to formamide by the enzyme cyanide hydratase. Such fungi can be immobilized and used in packed-cell columns to continuously detoxify cyanide. ICI Biological Products Business market a preparation of spray-dried fungal mycelia, 'CYCLEAR', to detoxify industrial wastes. Novo Industri have also introduced a cyanidase preparation to convert cyanide directly into formate and ammonia. Bacteria have been isolated that use cyanide as a source of nitrogen for growth. Because cyanide, as KCN or NaCN, is toxic for growth, the bacteria (Pseudomonas fluorescens) have to be grown in fed-batch culture with cyanide as the limiting nutrient. Cyanide is converted to carbon dioxide and ammonia (which is then assimilated) by an NADH-linked cyanide oxygenase system.

Bacteria↗

Encapsulation of proteins in biodegradable polymeric microparticles using electrospray in the Taylor cone-jet mode.

Solvent extraction (or evaporation from a W(1)/O/W(2)-dispersion), coacervation, and spray drying methods are commonly employed to encapsulate protein drugs in polymeric microparticles for sustained delivery applications. To overcome the limitations of these methods, a novel electrospray method was developed to encapsulate a model protein drug-bovine serum albumin (BSA) in biodegradable polymeric microparticles and examine the feasibility of the process in not denaturing the protein. Microparticles of approximately 20 microm diameter with corrugated surfaces and smooth surfaces were observed by scanning electron microscope. Confocal laser scanning microscope images showed that BSA was distributed evenly in microparticles. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was employed to investigate the protein integrity of BSA released from the polymer matrix after 38 days. No protein degradation was observed during the 38 days release. The secondary structure of released BSA was characterized by Fourier transform infrared (FTIR) and circular dichroism (CD), which suggested that the released BSA was almost identical to native BSA. The encapsulation efficiency could reach 76% by adjusting the amount of the additive Pluronic F127 and processing parameters. The release profile could be tailored by the fabrication process and the sustained release of BSA could endure for more than 1 month. More than 80% of the bioactivity of BSA (evaluated by BSA ELISA kit) could be maintained after releasing from polymer matrix. Findings of the present study demonstrate that this novel electrospray method is a promising approach to encapsulate bioactive materials such as proteins, enzymes, antibiotics, and DNA fragments in biodegradable polymeric particles.

Absorbable Implants↗

Effect of excipient composition on the biocompatibility of bupivacaine-containing microparticles at the sciatic nerve.

Microparticulate formulations are often used for experimental prolongation of nerve blockade. Here we examine the effect of excipient composition on the biocompatibility of bupivacaine-containing microparticles. Lipid-protein-sugar particles (LPSPs) composed of 3% (1.3-microm diameter) and 60% (4.7-microm diameter) (w/w) dipalmitoylphosphatidylcholine (DPPC) were produced by spray drying, containing 10% (w/w) bupivacaine. Rat sciatic nerve blocks with 75 mg of particles produced statistically similar durations of sensory nerve block [3% (w/w) DPPC particles: 301 min; 60% (w/w) DPPC particles: 321 min]. Examination of tissues 1 day after injection revealed large particle deposits and acute inflammation in animals that received 60% (w/w) DPPC particles. There were no visible deposits in those that received 3% (w/w) DPPC particles, and microscopic inflammation was reduced. The difference between groups was similar 4 days after injection. Two weeks after injection, there was no particulate mass in either group, and inflammation had largely resolved. In both groups, moderately severe myotoxicity was seen 1 and 4 days after injection but had largely resolved by 2 weeks. In summary, reduction in particles' DPPC content greatly improved biocompatibility without compromising duration of nerve blockade; the improvement was probably attributable to the enhanced rate of particle resorption.

Animals↗

Prolonged duration local anesthesia with lipid-protein-sugar particles containing bupivacaine and dexamethasone.

Glucocorticoids prolong block duration from polymeric microspheres containing bupivacaine, but not from unencapsulated drug. Here we investigate this effect applies to particles with much more rapid drug release and improved long-term biocompatibility. Male Sprague-Dawley rats were given sciatic nerve blocks with 75 mg of 3% or 60% (w/w) dipalmitoylphosphatidylcholine (DPPC) spray-dried lipid-protein-sugar particles (LPSPs) containing 10% (w/w) bupivacaine and 0%, 0.05%, or 0.1% (w/w) dexamethasone. Sensory nerve block from bupivacaine-containing 3% and 60% (w/w) DPPC particles without dexamethasone yielded blocks lasting 301 +/- 56 and 321 +/- 127 min, respectively. Addition of 0.05% (w/w) dexamethasone increased block durations to 610 +/- 182 and 538 +/- 222 min, respectively; increasing dexamethasone loading to 0.1% did not further increase duration. One day after injection, dexamethasone-containing particles resulted in lower inflammation scores and capsule thickness than dexamethasone-free particles, but the difference was gone by day 4. Excipient composition had prominent effects at all time points. For all groups, inflammation was largely resolved by 2 weeks after injection. Dexamethasone approximately doubled the duration of nerve block from bupivacaine-loaded LPSPs, while maintaining excellent biocompatibility. Such formulations could be useful in clinical applications when nerve blockade is needed for 24 hours or less.

Anesthesia, Local↗

Development of polymer film dosage forms of lidocaine for buccal administration: II. Comparison of preparation methods.

In previous studies, we prepared film dosage forms of lidocaine (LC) with hydroxypropylcellulose (HPC) as a film base using the solvent evaporation (SE) method. However, from the viewpoint of environmental issues, a reduction in organic solvent use in pharmaceutical and other industries is required. In this study, we prepared the LC films by direct compression of the physical mixture (DCPM method) and direct compression of the spray dried powder (DCSD method). Magnesium stearate, which was required as a lubricant for direct compression, showed no effect on the LC release rate. The LC release rate (%/h) was independent of the compression pressure, but a higher pressure was preferable to easily remove the film from the punches. An increase in the film weight decreased the LC release rate expressed in %/h, whereas no significant effect of film weight was observed on the LC release rate from unit surface area expressed in mg/h/cm(2). The LC release rate (%/h) was independent of the LC content, suggesting that the LC release rate (mg/h) can be quantitatively controlled by changing the LC content in the formulation. The LC release rate and penetration rate were affected by the preparation method; that is, DCPM method < DCSD method < SE method. The LC penetration rates through excised hamster oral mucosa were linearly correlated to the release rate of un-ionized LC, which was estimated by the LC release rate multiplied by the un-ionized fraction of LC for the HPC film dosage form.

Administration, Buccal↗

An experimental divalent vaccine based on biodegradable microspheres induces protective immunity against tetanus and diphtheria.

In an endeavor towards development of multivalent vaccines based on biodegradable microspheres, we tested the immunologic performance of several divalent microsphere formulations against tetanus and diphtheria. Microspheres were made by separate microencapsulation of tetanus and diphtheria toxoid in poly(lactide-co-glycolide) by either spray-drying or coacervation. Guinea pigs were subcutaneously immunized by a single injection of the divalent vaccines or, for control, an equivalent dose of a licensed vaccine containing both antigens adsorbed on aluminium hydroxide. All microsphere formulations were strongly immunogenic, irrespective of particle size and hydrophobicity. End point titers of ELISA antibodies, mainly of the IgG1 subtype, were comparable to those obtained after immunization with the licensed vaccine. The microspheres provided increasing levels of antibodies, during the 16 weeks of testing, and the antibodies were weakly polarized towards tetanus. The induced antibodies were also toxin neutralizing, as determined for both diphtheria (1-4 IU/mL) and tetanus (5-9 IU/mL) 8 weeks after immunization. These neutralization levels were several orders of magnitude above the level considered minimum for protection (0.01 IU/mL). When the animals were challenged with tetanus or diphtheria toxins 6 weeks after immunization, microsphere vaccines produced protective immunity that was comparable to or better than that induced by the licensed divalent vaccine. In conclusion, this study showed that a single administration of biodegradable microsphere vaccines provided protective immunity against diphtheria and tetanus, and that this immunization approach might be feasible for multivalent vaccines.

Animals↗

Enhancing the oral bioavailability of the poorly soluble drug dicumarol with a bioadhesive polymer.

This article investigates the effect of particle size and the incorporation of a bioadhesive polymer, poly(fumaric-co-sebacic) anhydride p(FA:SA), on the relative bioavailability of dicumarol. A novel method was used to reduce particle size of the drug, and encapsulated formulations were fabricated using a phase inversion technique to produce nanospheres and microspheres with varying size. Groups of Yorkshire swine were catheterized and gavaged after fasting for 12 h with each formulation in a 50 mg/mL suspension. Blood was collected at different time points, from 0 to 96 h, and pharmacokinetic analysis revealed that formulations incorporating the smaller drug particles showed the highest bioavailability: micronized drug with 7% p(FA:SA) 17:83 polymer had 190% relative bioavailability, and phase inverted p(FA:SA) 17:83 microspheres with 31% (w/w) loading had 198% relative bioavailability to spray dried formulation. Formulations with larger drug particles achieved 71% relative bioavailability. A nonadhesive formulation, fabricated with poly(lactic acid) (PLA), showed 91% relative bioavailability. Both particle size and polymer composition play a role in oral absorption of dicumarol.

Adhesives↗

The influence of excipients on drug release from hydroxypropyl methylcellulose matrices.

The influence of commonly used excipients, spray-dried lactose (SDL), microcrystalline cellulose (MCC), and partially pregelatinized maize starch (Starch 1500) on drug release from hydroxypropyl methylcellulose (HPMC, hypromellose) matrix system has been investigated. A model formulation contained 30%w/w drug, 20%w/w HPMC, 0.5%w/w fumed silica, 0.25%w/w magnesium stearate, and 49.25%w/w filler. Chlorpheniramine maleate and theophylline were used as freely (1 in 4) and slightly (1 in 120) water-soluble drugs, respectively. It was found that for both drugs, addition of 20 to 49.25%w/w Starch 1500 resulted in a significant reduction in drug release rates compared to when MCC or SDL was used. The study showed that using lactose or microcrystalline cellulose in the formulations resulted in faster drug release profiles. Partially pregelatinized maize starch contributed to retardation of both soluble and slightly soluble drugs. This effect may be imparted through synergistic interactions between Starch 1500 and HPMC and the filler actively forming an integral part within the HPMC gel structure.

Chemistry, Pharmaceutical↗

Effect of amino acids on the dispersion of disodium cromoglycate powders.

Modified disodium cromoglycate powders were prepared by co-spray drying with different concentrations of leucine, phenylalanine, tryptophan, methionine, asparagine, and arginine. Amorphous spherical particles of the same size and density where obtained which, however, exhibited different surface properties as measured by the inverse gas chromatography (IGC) and X-ray photoelectron spectroscopy (XPS) techniques. The surface energy parameters, such as the dispersive component of surface free energy of the sample, gammaSD, and the total solubility parameter, delta, were significantly lower in the presence of nonpolar chain amino acids, particularly with leucine and phenylalanine, than pure DSCG. However no quantitative relationship between these parameters, the additive concentrations, and the fine particle fractions, FPF, determined for different inhalers and air flow rates, was observed. The FPF significantly increased with addition of leucine and this effect was attributed to reduced intermolecular interactions between leucine and disodium cromoglycate molecules, as indicated by the difference in corresponding Hansen solubility parameters. Decrease of interparticle interactions for leucine-containing powders also led to a lesser dependence of FPF on the flow rate and inhaler type.

Aerosols↗

Prediction of tablet hardness based on near infrared spectra of raw mixed powders by chemometrics.

The purpose of this research is to elucidate the effect of lubricant mixing on tablet hardness by near-infrared (NIR) chemometrics as a basic study of process analytical technology. Formulation cellulose (F-C) consisted of sulpyrine (SP), microcrystalline cellulose (MC), and magnesium stearate (MgSt). Formulation lactose/starch (F-L) consisted of SP bulk drug powder, spray-dried lactose (SL), corn starch (CS), and MgSt. First, F-L and F-C without MgSt were mixed in a twin-shell mixer for 60 min. MgSt was added to the mixed powder, and was mixed for various mixing times, after which the mixed powders were compressed by 8-mm diameter punch and die. NIR spectra of raw mixed powders of F-L and F-C were taken using a reflection type of Fourier transform NIR spectra spectrometer, and chemometric analysis was performed using principal component regression (PCR). The tablet hardnesses of F-L and F-C decreased with increasing mixing time. All NIR spectra of the mixed powders of F-L and F-C fluctuated depending on mixing time. In order to predict tablet hardness before tablet compression, NIR spectra of F-L and F-C mixed powders were analyzed and evaluated for hardness by PCR. The minimum standard error of cross-validation values could be realized by using five- and six-principal component models, respectively. In the cases of F-L and F-C, the relationships between the actual and predicted tablet hardnesses showed straight lines, respectively. In the regression vectors of F-L and FC, the peaks related to hydrogen groups of SP, CS, and MC appeared as positive peaks. In contrast, the peaks related to hydrocarbon due to MgSt appeared as negative peaks in the regression vectors. The calibration models to evaluate the tablet hardness were obtained based on NIR spectra of raw mixed powders by PCR. This approach to predicting tablet hardness prior to compression could be used as a routine test to indicate the quality of the final product without spending time and energy to produce samples of questionable quality.

Cellulose↗