Application of hydroxypropyl methylcellulose as a protective agent against magnesium stearate induced crystallization of amorphous itraconazole.

Itraconazole is a fungicide drug which has low bioavailability due to its poor water solubility. Amorphous solid dispersion (ASD) is a tool that has the potential to greatly increase the dissolution rate and extent of compounds. In this work, the dissolution of tablets containing the ASD of itraconazole with either hydroxypropyl methylcellulose (HPMC) or vinylpyrrolidone-vinyl acetate copolymer (PVPVA) was compared in order to find a formulation which can prevent the drug from the precipitation caused by magnesium stearate. Formulations containing the PVPVA-based ASD with HPMC included in various forms could reach 90% dissolution in 2 h, while HPMC-based ASDs could release 100% of the drug. However, HPMC-based ASD had remarkably poor grindability and low bulk density, which limited its processability and applicability. The latter issue could be resolved by roller compacting the ASD, which significantly increases the bulk density and the flowability of the powder blends used for tableting. This roller compaction step might be a base for the industrial application of HPMC-based, electrospun ASDs.

Investigation of Deteriorated Dissolution of Amorphous Itraconazole: Description of Incompatibility with Magnesium Stearate and Possible Solutions.

Disadvantageous crystallization phenomenon of amorphous itraconazole (ITR) occurring in the course of dissolution process was investigated in this work. A perfectly amorphous form (solid dispersion) of the drug was generated by the electroblowing method (with vinylpyrrolidone-vinyl acetate copolymer), and the obtained fibers were formulated into tablets. Incomplete dissolution of the tablets was noticed under the circumstances of the standard dissolution test, after which a precipitated material could be filtered. The filtrate consisted of ITR and stearic acid since no magnesium content was detectable in it. In parallel with dissolution, ITR forms an insoluble associate, stabilized by hydrogen bonding, with stearic acid deriving from magnesium stearate. This is why dissolution curves do not have the plateaus at 100%. Two ways are viable to tackle this issue: change the lubricant (with sodium stearyl fumarate >95% dissolution can be accomplished) or alter the polymer in the solid dispersion to a type being able to form hydrogen bonds with ITR (e.g., hydroxypropyl methylcellulose). This work draws attention to one possible phenomenon that can lead to a deterioration of originally good dissolution of an amorphous solid dispersion.

Detailed stability investigation of amorphous solid dispersions prepared by single-needle and high speed electrospinning.

In this research the long-term stability (one year) of amorphous solid dispersions (ASDs) prepared by high speed electrospinning was investigated at 25 °C/60% relative humidity (RH) (closed conditions) and 40 °C/75% RH (open conditions). Single needle electrospinning and film casting were applied as reference technologies. Itraconazole (ITR) was used as the model API in 40% concentration and the ASDs consisted of either one of the following polymers as a comparison: polyvinylpyrrolidone-vinyl acetate 6:4 copolymer (no hydrogen bonds between API and polymer) and hydroxypropyl methylcellulose (possible hydrogen bonds between oxo or tertiary nitrogen function of API and hydroxyl moiety of polymer). DSC, XRPD and dissolution characteristics of samples at 0, 3 and 12 months were investigated. In addition, Raman maps of certain electrospun ASDs were assessed to investigate crystallinity. A new chemometric method, based on Multivariate Curve Resolution-Alternating Least Squares algorithm, was developed to calculate the spectrum of amorphous ITR in the matrices and to determine the crystalline/amorphous ratio of aged samples. As it was expected ITR in single needle electrospun SDs was totally amorphous at the beginning, in addition hydroxypropyl methylcellulose could keep ITR in this form at 40 °C/75% RH up to one year due to the hydrogen bonds and high glass transition temperature of the SD. In polyvinylpyrrolidone-vinyl acetate matrix ITR remained amorphous at 25 °C/60% RH throughout one year. Materials prepared by scaled-up, high throughput version of electrospinning, which is compatible with pharmaceutical industry, also gained the same quality. Therefore these ASDs are industrially applicable and with an appropriate downstream process it would be possible to bring them to the market.

Downstream processing of polymer-based amorphous solid dispersions to generate tablet formulations.

Application of amorphous solid dispersions (ASDs) is considered one of the most promising approaches to increase the dissolution rate and extent of bioavailability of poorly water soluble drugs. Such intervention is often required for new drug candidates in that enablement, bioavailability is not sufficient to generate a useful product. Importantly, tableting of ASDs is often complicated by a number of pharmaceutical and technological challenges including poor flowability and compressibility of the powders, compression-induced phase changes or phase separation and slow disintegration due to the formation of a gelling polymer network (GPN). The design principles of an ASD-based system include its ability to generate supersaturated systems of the drug of interest during dissolution. These metastable solutions can be prone to precipitation and crystallization reducing the biopharmaceutical performance of the dosage form. The main aim of the research in this area is to maintain the supersaturated state and optimally enhance bioavailability, meaning that crystallization should be delayed or inhibited during dissolution, as well as in solid phase (e.g., during manufacturing and storage). Based on the expanding use of ASD technology as well as their downstream processing, there is an acute need to summarize the results achieved to this point to better understand progress and future risks. The aim of this review is to focus on the conversion of ASDs into tablets highlighting results from various viewpoints.

Effective dose of miners due to natural radioactivity in a manganese mine in Hungary.

In this study, short-term radon (RnDP) and thoron (TnDP) progeny measurements and dose estimation were carried out in winter and summer in a manganese mine, Hungary. Gamma-ray dose rate originating from external sources and (222)Rn and (226)Ra contents of spring-water from a mine was also measured. During working hours RnDP and TnDP concentration values changed between 12.1-175 and 0.14-0.42 Bq m(-3), respectively. The (222)Rn and (226)Ra concentration values in the karst spring-water were ∼6 Bq dm(-3) and 16 mBq dm(-3), respectively. The radiation dose resulting from the consumption of karst spring-water was negligible. The doses from the inhalation of TnDP and external gamma radiation were of the same magnitude, ∼0.1 mSv y(-1), which was rather negligible related to the estimated radiation dose of 5 mSv y(-1) from RnDP.

Radon and thoron parallel measurements in Hungary.

Hungarian detectors modified and developed at the National Institute of Radiological Sciences (NIRS), Japan were placed at different sites, including homes and underground workplaces in Hungary, in order to gain information on the average radon (222Rn) and thoron (220Rn) concentration levels. Measurements were carried out in dwellings in a village and a manganese mine in Hungary. The radon and thoron concentrations in the dwellings of the village in the summer period were found to be 154 (17-1083) and 98 (1-714) Bq m(-3), respectively. Considering the results of other radon measurements during the winter (814 Bq m(-3)) and summer (182 Bq m(-3)) periods, the thoron concentrations were also expected to be higher in winter. In the manganese mine, radon and thoron were measured at 20 points for 6 months, changing the detectors each month. The averages were 924 (308-1639) and 221 (61-510) Bq m(-3) for radon and thoron, respectively. These results showed significant variance with the date and place of the measurement.

Evaluation and clinical application of a new method for measuring activity of von Willebrand factor-cleaving metalloprotease (ADAMTS13).

Thrombotic thrombocytopenic purpura (TTP) is associated with acquired or congenital deficiency of a plasma von Willebrand factor-cleaving protease (VWFcp). Based on partial amino acid sequence and genome-wide linkage analysis of pedigrees with congenital TTP, VWFcp was recently identified as a new member of the ADAMTS family and designated ADAMTS13. We developed a new, rapid, and simple method for measuring VWFcp activity based on the positive correlation between VWF multimeric size and Ristocetin cofactor activity (VWF:RCo). After dilution of plasma with low ionic Tris buffer and activation of the protease with barium chloride, a VWF concentrate is digested in the presence of urea. Subsequently, the residual VWF:RCo of the samples is assessed and used to calculate the VWFcp activity of the samples. The accuracy of the new technique is verified by estimating VWFcp activity for 282 plasma samples with the RCo-based assay and the original immunoblotting assay. The method is reproducible as shown by low intra- and interassay coefficients of variation (2.8% and 7.5% for normal samples, respectively, and 8.7% and 12.9% for abnormal samples, respectively). Furthermore, the clinical application of the new method is illustrated by measuring VWFcp of 14 patients with 22 episodes of acute TTP as well as other thrombotic, thrombocytopenic, or hemolytic disorders. Severe VWFcp deficiency was restricted to patients with acute, classic TTP. The majority of patients with low titer inhibitor respond to plasma exchange treatment with increase of VWFcp activity, whereas VWFcp deficiency persists in some patients with high titer inhibitor despite clinical remission.

High doses of methylene blue/light treatment crosslink the A-alpha-subunit of fibrinogen: influence of this photooxidization on fibrinogen binding to platelets.

Irradiation in the presence of 1 microM methylene blue (MB) inactivates enveloped viruses in human plasma. This method is a useful tool to enhance the safety of human fresh-frozen plasma in blood donor banks. Via a photooxidative mechanism, viral structures as well as plasma proteins, especially fibrinogen, can be damaged. We investigated the effect of MB/light-induced photooxidative modification on the structure of fibrinogen and its function in the interaction with platelets in the concentration range of 150 microM MB. Densitometric scanning of reduced SDS-PAGE showed a decrease in the Aalpha-subunit of fibrinogen as well as an increase in high-molecular-weight (HMW) aggregates with increasing MB concentrations. The HMW aggregates may be due to covalent crosslinking of single Aalpha-subunits via photooxidation induced by MB/light treatment. Fibrinogen treated with high doses of MB (50 microM) showed a weaker binding to the fibrinogen receptor (GP IIb/IIIa) on the platelet surface and a decrease in platelet aggregation after stimulation with ADP and photooxidized fibrinogen. These effects were not detected in fibrinogen isolated from virus-inactivated plasma (1 microM MB). This was in agreement with normal binding of fibrinogen to GP IIb/IIIa on platelets.

The Thrombostat 4000. A sensitive screening test for von Willebrand's disease.

The determination of the bleeding time (BT) is an essential diagnostic tool for von Willebrand's disease (vWD). However, the standardized Simplate BT still displays many variables and disadvantages. The present study reports on the sensitivity of the in vitro bleeding test (IVBT) in 51 vWD cases of different types and severity in comparison to the Simplate BT, and the correlation of both to each other as well as to von Willebrand factor (vWF:RCo) activity. The IVBT was performed in two modifications (2 mmol/L CaCl2 and 4 mmol/L ADP) on the Thrombostat 4000. The IVBT, particularly with CaCl2, showed clearly higher sensitivity than the BT (CaCl2: 84.3%, ADP: 61.7%, CaCl2 + ADP: 86.3, BT: 52.9%). The BT even failed in one patient with severe (type 2B) and in three with moderate vWD. The IVBT only failed in very mild forms of the disease (vW-F:RCo > 25%). In addition, the IVBT with ADP showed a close correlation to the vWF:RCo activity (r2 = 0.73). The significantly lower correlation of the BT with vWF:RCo (r2 = 0.49) was particularly due to the poor results in vWD of type 2 (type 2:r2 = 0.29; types 1 and 3:r2 = 0.61). Finally, BT and IVBT-ADP correlated with each other (r2 = 0.53), a rather good correlation considering that both are complex functional tests. It can be concluded from our study that the IVBT not only may replace the BT for most applications, but is clearly superior to BT for the screening (IVBT-CaCl2) and control of therapy in vWD.

Experience with Haemate P in von Willebrand's disease in adults.

The virally inactivated pasteurized FVIII concentrate Haemate P contains nearly intact vWF multimers. It is currently the treatment of choice to achieve satisfactory hemostasis for moderate to severe vWD and for patients with variants of vWD that cannot be adequately treated with DDAVP or for whom DDAVP is contraindicated. Therefore, we treated patients with type Ia, type IIa, type IIb and type III vWD with Haemate P. A correction of the hemostatic defect was seen in all patients. The type of bleeding events included 24 gastrointestinal, 18 other mucosal, 5 central nervous system, 15 orthopedic and 4 other. 28 dental surgical procedures, 9 operative deliveries, 5 tonsillectomies, 17 orthopedic and 11 miscellaneous surgeries were performed under the cover of Haemate P.