Development of gel-forming lyophilized formulation with recombinant human thrombin

The objective of this work was development and evaluation of gel-forming lyophilized formulation with recombinant human thrombin for topical administration. The influence of pH, ionic strength and buffer type on protein stability was evaluated as part of the pre-formulation screening studies. Results indicated an optimal pH from 6.0 to 7.0 and increased stability with increasing content of sodium chloride. The tested buffer types had no significant effect on thrombin stability. For further development, thermosensitive Pluronic® F-127 was employed as a bulking and gelling agent. Physical and mechanical characterization and viscosity measurement confirmed the gel-forming properties of the formulation at the application temperature of 32?°C. Several techniques (addition of well-soluble polyols, different freezing protocols and reconstitution under vacuum) were tested to decrease the reconstitution time. The obtained results revealed that a vacuum in the vial headspace is crucial for acceptable reconstitution. The freeze drying process has no negative impact on recombinant thrombin stability, and this was confirmed by reverse-phase-HPLC, activity assay and optical density measurements.     

Freeze drying optimization of polymeric nanoparticles for ocular flurbiprofen delivery: effect of protectant agents and critical process parameters on long-term stability

Context: The stabilization of flurbiprofen loaded poly-?-caprolactone nanoparticles (FB-P?CL-NPs) for ocular delivery under accurate freeze-drying (FD) process provides the basis for a large-scale production and its commercial development.

Objective: Optimization of the FD to improve long-term stability of ocular administration’s FB-P?CL-NPs.

Methods: FB-P?CL-NPs were prepared by solvent displacement method with poloxamer 188 (P188) as stabilizer. Freezing and primary drying (PD) were studied and optimized through freeze-thawing test and FD microscopy. Design of experiments was used to accurate secondary drying (SD) conditions and components concentration. Formulations were selected according to desired physicochemical properties. Furthermore, differential scanning calorimetry (DSC) and X-ray diffraction (XRD) were used to study interactions components.

Results: Optimized FB-P?CL-NPs, stabilized with 3.5% (w/w) P188 and protected with 8% (w/w) poly(ethylene glycol), was submitted to precooling at +10?°C for 1?h, freezing at ?50?°C for 4?h, PD at +5?°C and 0.140 mbar for 24?h and a SD at +45?°C during 10?h. These conditions showed 188.4?±?1.3?nm, 0.087?±?0.014, 85.5?±?1.4%, 0.61?±?0.12%, ?16.4?±?0.1?mV and 325?±?7 mOsm/kg of average size, polydispersity index, entrapment efficiency, residual moisture, surface charge and osmolality, respectively. It performed a long-term stability >12 months. DSC and XRD spectra confirmed adequate chemical interaction between formulation components and showed a semi-crystalline state after FD.

Conclusions: An optimal freeze dried ocular formulation was achieved. Evidently, the successful design of this promising colloidal system resulted from rational cooperation between a good formulation and the right conditions in the FD process.     

Assessment of Degree of Disorder (Amorphicity) of Lyophilized Formulations of Growth Hormone Using Isothermal Microcalorimetry

When determining the degree of disorder of a lyophilized cake of a protein, it is important to use an appropriate analytical technique. Differential scanning calorimetry (DSC) and X‐ray powder diffraction (XRPD) are the most commonly used thermo‐analytical techniques for characterizing freeze‐dried protein formulations. Unfortunately, these methods are unable to detect solid‐state disorder at levels < 10%. Also, interpretation of DSC results for freeze‐dried protein formulations can be difficult, as a result of the more complex thermal events occurring with this technique. For example, proteins can inhibit the thermally induced recrystallization of the lyophilized cake, resulting in potential misinterpretation of DSC degree of disorder results. The aim of this investigation was to study the use of isothermal microcalorimetry (IMC) in the assessment of degree of solid‐state disorder (amorphicity) of lyophilized formulations of proteins. For this purpose, two formulations of growth hormone were prepared by lyophilization. These formulations consisted of the same amounts of protein, mannitol, glycine, and phosphate buffer, but differed in the freeze‐drying procedure. After lyophilization, the recrystallization of the samples was studied using IMC at 25°C under different relative humidities (58–75%). The effect of available surface area was studied by determining the heat of recrystallization (Q) of the samples before and after disintegration of the cakes. The results showed that, in contrast to DSC, IMC allowed detection of the recrystallization event in the formulations. Although both formulations were completely disordered and indistinguishable according to XRPD method, IMC revealed that formulation B had a different solid‐sate structure than formulation A. This difference was the result of differences in the freeze‐drying parameters, demonstrating the importance of choosing appropriate analytical methodology.     

Synthesis and characterization of sol-gel derived TiO<Subscript>2</Subscript> thin films: Effect of different pretreatment process

Titanium dioxide (TiO₂) thin films have been successfully synthesized deposited on glass substrates by the sol-gel dip-coating method through different pretreating processes, including heated at 100, 500°C, via freeze drying, microwave heating for 10 min and subsequently annealed at 500°C for 2 h. The as-synthesized TiO₂ films were characterized using X-ray diffraction (XRD), atomic force microscopy (AFM) and ultravioletvisible (UV-vis) absorption spectra analysis technology. The preparation of the precursor sols and TiO₂ films were described in detail. Effects of 100, 500°C, freeze drying and microwave heating pretreatment on crystalline structure, surface morphology, roughness, particle size, optical property and electronic transition of TiO₂ thin films have been primarily investigated. The XRD results demonstrate that the TiO₂ films were well-crystallized and consisted of anatase TiO₂ phase only with (101) plane. The average crystalline size is only about 15 nm at 100°C pretreatment and the absorption edge shifts to shorter wavelength comparing with that at 500°C, freeze drying and microwave heating pretreatment. Pretreatment process is important during the preparation of thin films and has obviously effect on the structure and optical property of TiO₂ films due to the different heating mechanisms.     

Comparison of Co�CC and Pd�CC Eutectic-Point Cells for Thermocouple Calibration Between LNE-Cnam and NMIJ

Evaluation of the melting temperatures of two Co?CC and two Pd?CC eutectic-point cells were performed using three Pt/Pd thermocouples constructed at LNE-Cnam and NMIJ. During the comparison of cells, Pt/Pd thermocouples were evaluated at the Ag fixed point, at which their drifts and inhomogeneity represented differences within 0.03 °C and 0.14 °C, respectively. One Co?CC cell and one Pd?CC cell were designed and constructed at LNE-Cnam, while one Co?CC cell and one Pd?CC cell were designed and constructed at NMIJ. In spite of differences in cell design and in materials sources, melting points of Co?CC and Pd?CC eutectic-point cells realized using LNE-Cnam high-temperature furnace agreed within approximately 0.02 °C and 0.01 °C, respectively. Furthermore, the realizations of the Pd?CC eutectic point at LNE-Cnam and NMIJ agreed within a temperature equivalent of approximately 0.11 °C. The uncertainty of the realization was estimated to be approximately 0.14 °C (k = 2), with a major contribution from the inhomogeneity of the thermocouple.     

Cobalt�CCarbon Eutectic Fixed Point for Contact Thermometry

Two Co?CC eutectic fixed points were constructed for thermocouple calibration. The eutectic fixed points were measured with a Pt/Pd thermocouple calibrated at the freezing temperatures of Sn, Zn, Al, Ag, and Au. A temperature of (1323.99 ± 0.52) °C (k = 2) was determined via this method. The cell design allowed filling to be accomplished in a single step. Each cell was held above 1300 °C for at least 42 h and was subjected to at least 20 melt/freeze cycles with no mechanical failure occurring.     

Stability of 10 mg/mL cefuroxime solution for intracameral injection in commonly used polypropylene syringes and new ready-to-use cyclic olefin copolymer sterile vials using the LC-UV stability-indicating method

Context: Injecting intracameral cefuroxime has been found beneficial in reducing the risk of postoperative endophthalmitis but its use has been limited through a lack of approved marketing and of ready-to-use single-units as well as the problem of aseptic compounding.

Objective: Our aim was to assess a new automated primary packaging system which should ensure a higher level of sterility, thanks to its closed, sterile, ready-to-use polymer vial called “Crystal® vial”. The chemical stability of a 10?mg/mL cefuroxime solution was compared in 1?mL Crystal® vials and 1?mL Luer-lock polypropylene syringes (actual reference) to eliminate any potential and specific interactions with its cyclic olefin copolymer (COC) body and elastomer stopper.

Methods: Cefuroxime solution was introduced into vials and syringes and stored at ?20?°C, +5?°C and +25°C/60% Relative Humidity. Cefuroxime concentration and the relative amount of the main degradation product (descarbamoyl-cefuroxime) were both determined by an HPLC/UV method indicating stability. Solutions were considered steady if the concentration remained at over 90% of the initial value. In the adapted storage conditions, the evolution of osmolality, pH and sterility was assessed.

Results: Stability profiles were identical between vials and syringes in all storage and temperature conditions. The solution was stable (cefuroxime concentration, pH and osmolality) and still sterile for 365 days at ?20°C. The concentration fell below 90% after 21 days at +5?°C and after 16?h at +25°C/60%s relative humidity.

Conclusions: The COC and thermoplastic elastomer of the vials had no impact on the degradation process confirming its possible use for a ready-to-use cefuroxime solution single-unit dose.     

Comparison of Re�CC Fixed-Point Cells and Their T90 Temperatures Between NMIJ and VNIIOFI

The comparison of Re?CC fixed-point cells built at NMIJ and VNIIOFI was carried out. Both cells were built using 5N purity rhenium of the same supplier, and the crucibles had similar dimensions. The cells were heated in the same furnace and measured on consecutive days using a radiation thermometer. To eliminate the effect of possible instability of the thermometer, a third Re?CC cell was used as a reference: it was placed in another furnace and was measured each day just after the cell to be compared. The melting-temperature difference of the NMIJ cell to the VNIIOFI cell was 0.045 °C with a standard uncertainty of 0.033 °C. In the course of this comparison, a comparison of the temperature scales between the NMIJ and the VNIIOFI at the Re?CC point (2475 °C) was performed using the NMIJ Re?CC cells as transfer standards. The difference between the NMIJ and the VNIIOFI scales was found to be ?0.8 °C, which was within the combined uncertainty of 1.4 °C (k = 2).     

Pharmaceutical development of an amorphous solid dispersion formulation of elacridar hydrochloride for proof-of-concept clinical studies

Objective: A novel tablet formulation containing an amorphous solid dispersion (ASD) of elacridar hydrochloride was developed with the purpose to resolve the drug’s low solubility in water and to conduct proof-of-concept clinical studies.

Significance: Elacridar is highly demanded for proof-of-concept clinical trials that study the drug’s suitability to boost brain penetration and bioavailability of numerous anticancer agents. Previously, clinical trials with elacridar were performed with a tablet containing elacridar hydrochloride. However, this tablet formulation resulted in poor and unpredictable absorption which was caused by the low aqueous solubility of elacridar hydrochloride.

Methods: Twenty four different ASDs were produced and dissolution was compared to crystalline elacridar hydrochloride and a crystalline physical mixture. The formulation with highest dissolution was characterized for amorphicity. Subsequently, a tablet was developed and monitored for chemical/physical stability for 12 months at +15–25?°C, +2–8?°C and ?20?°C.

Results: The ASD powder was composed of freeze dried elacridar hydrochloride–povidone K30–sodium dodecyl sulfate (1:6:1, w/w/w), appeared fully amorphous and resulted in complete dissolution whereas crystalline elacridar hydrochloride resulted in only 1% dissolution. The ASD tablets contained 25?mg elacridar hydrochloride and were stable for at least 12 months at –20?°C.

Conclusions: The ASD tablet was considered feasible for proof-of-concept clinical studies and is now used as such.     

阴沟肠杆菌定量标准物质的研制

采用真空冷冻干燥法和滤膜法定值技术,研制了球状的阴沟肠杆菌定量标准物质。经保护剂配方及冻干程序参数优化后,阴沟肠杆菌的平均存活率达51%,形状均匀一致、外观光滑无塌陷,能完全复水速溶。通过结晶紫中性胆盐琼脂培养基平板计数法考察均匀性和稳定性,证明该标准物质均匀性良好,-80 ℃至少可稳定保存6个月。该标准物质由6家实验室采用滤膜法联合定值,经不确定度评定,标称值为（2.0±0.1）log10 (cfu/瓶),相对扩展不确定度为10%(k=2)。标准物质中阴沟肠杆菌活菌数含量水平为10² cfu/瓶,可以直接溶解使用,无需稀释,使用方便,同时减少了测量不确定度来源。     

Effect of modified atmosphere packaging and superchilled storage on the shelf‐life of farmed ready‐to‐cook spotted wolf‐fish (Anarhichas minor)

Modified atmosphere packaging (MAP) combined with superchilling (?1°C) was evaluated as a mild preservation method for farmed spotted wolf‐fish (Anarhichas minor). Portions were packaged in air and in CO2:N2 (60%:40%) atmosphere with a gas:product ratio of approximately 1, at superchilled (?1.0°C ± 0.2°C) or chilled (+4.0°C ± 0.2°C) temperatures. A reduced bacterial growth (p < 0.001), measured as aerobic plate counts (APC) and psychrotrophic bacteria, was observed in modified atmosphere (MA) packaged wolf‐fish at both ?1°C and +4°C, compared to portions in air. MA‐packaged wolf‐fish had improved odour and flavour scores (p < 0.05), but also a higher drip loss than fish stored in air. A shelf‐life of 15 days was obtained at ?1°C for MA‐packaged fish compared to 8–10 days in air, and at 4°C the shelf‐life was 13 days in MA and 6–8 days in air. Copyright © 2006 John Wiley & Sons, Ltd.     

A Four-Component Study for Estimating Solubilities of a Poorly Soluble Compound in Multisolvent Systems Using a Scheffé-Type Model

Abstract

The equilibrium solubility of GF1209184, a poorly soluble compound with potential use as a multidrug resistance (MDR) inhibitor of P-glycoprotein, was studied at 25°C in multisolvent systems containing polyethylene glycol 300, polysorbate 80, ethanol, and water. The objective was to determine the feasibility, with respect to solubility, of formulating a concentrated formulation for product presentation in an ampule or vial. Data were fit to a quadratic Scheffé-type model with excellent correlation between the experimentally determined and fitted equilibrium solubilities (R² = 0.9875, slope = 1.043). Solubilities greater than 4 mg base/mL at 25°C were determined for mixtures in this study, making it feasible, with regard to solubility, to formulate a concentrated vial or ampule formulation. Maximum solubility, however, was dependent on the ability to adjust the apparent pH to ≤3.5 in the cosolvent/surfactant systems studied.     

Thermal stability of nanocrystalline CeO<Subscript>2</Subscript> prepared through freeze drying

The growth of ceria nanoparticles during annealing at high temperatures (up to 700°C) has been studied by X-ray diffraction, transmission electron microscopy, and low-temperature nitrogen adsorption measurements. The results indicate that CeO₂ nanopowders prepared using freeze drying offer enhanced thermal stability.     

Application of salt engineering to reduce/mask bitter taste of clindamycin

Abstract

Palatability of a formulation is one of the primary requirements for therapeutic compliance in children. Clindamycin (CLN) often prescribed to children to treat various infections. However, it has a bitter taste and bad smell. The focus of the present investigation was to develop salt of CLN with a commonly used sweetener such as cyclamic acid (CYA) to improve the palatability. The salt forms were prepared by solubilization crystallization method and characterized by Fourier transformed infrared (FTIR), Near infrared (NIR), Raman, X-ray powder diffraction, scanning electron microscopy, solubility, dissolution, and solid-state physical and chemical stability at 25?°C/60% RH and 40?°C/75% RH for 1 month and 60?°C for 2 weeks. Spectroscopic and diffraction data indicated the formation of a new solid phase, which was different from hydrochloride salt of CLN. Shape of crystal was rectangular prism. Stoichiometric ratio between CLN and CYA in the new salt CLN-CYA was 1:1 and its melting point was 85.6?°C. There was a 2.4-fold reduction in solubility of CLN-CYA at pH 4 compared with CLN-HCl. Moreover, the dissolution rate and extent were similar between the two salts and meeting USP requirement of 85% dissolution in 30?min. Salt was physically and chemically stable at 60?°C, 25?°C/60% RH, and 40?°C/75% RH conditions but hygroscopic at high humidity condition. In conclusion, new salt will provide a new avenue for the development of a palatable formulation of CLN.     

The Stability of Insulin in Solid Formulations Containing Melezitose and Starch. Effects of Processing and Excipients

ABSTRACT

Solid insulin formulations obtained by different methods of preparation were compared with respect to chemical stability and morphology. Spray- and freeze-drying, solution enhanced dispersion by supercritical fluids (SEDS) and precipitation into starch microspheres were the methods used for preparation of solid powders. The excipients applied were melezitose, starch, and sodium taurocholate. The stability of the samples was evaluated after storage in open containers at 25°C and 30% RH for 6 months.

All samples were amorphous after processing and storage as detected by XRD, except for the starch microspheres which were semi-crystalline. The spray- and freeze-dried samples containing melezitose and sodium taurocholate experienced a significant water uptake during storage, resulting in changes in morphology and disappearance of Tg. However, the chemical stability of these samples did not seem to be affected by the water uptake. Changes in morphology were not observed for the SEDS powders and the starch microspheres.

The chemical stability of the samples was assessed by HPLC. In general, conventional spray- and freeze drying resulted in samples with higher chemical stability compared to SEDS powders and starch microspheres. Nevertheless, the excipients applied were observed to be of major importance, and further optimization of the formulation as well as processing conditions may lead to slightly different conclusions.     

Stability of Tungsten�CRhenium Thermocouples in the Range from 0?��C to 1500?��C

The effect of exposure up to 1500 °C on emf values of type C (95 % tungsten 5 % rhenium vs. 74 % tungsten 26 % rhenium) thermocouples were evaluated. Three thermocouples consisting of thermocouple wires of 0.5 mm diameter, twin-bore beryllia tubes, and tantalum sheaths were prepared. After three type C thermocouples were calibrated in the range from 0 °C to 1550 °C, which confirmed insignificant difference among them, the drifts of two among them were measured at the palladium?Ccarbon (Pd?CC) eutectic point (1492 °C). They indicated a similar tendency, where the emf of thermocouples increased rapidly within the first 30 h, and after that, decreased gradually. To investigate the mechanism of the drift, the inhomogeneities of thermocouples were examined at 160 °C using a water heat-pipe furnace during the drift measurements at the Pd?CC eutectic point. It was found that the increase of emf within the first 30 h exposure at around 1500 °C was caused by the emf change due to inhomogeneity above 700 °C, and after that, the decrease of emf was caused by that around 1400 °C.     

Melting Temperature of High-Temperature Fixed Points for Thermocouple Calibrations

Thermocouples can be calibrated at pure metal ingot-based fixed points at temperatures up to the freezing point of copper (1084.62 °C). For Pt/Pd thermocouples, the deviation from the accepted reference function very often takes an approximately linear form up to the copper fixed point. The calibration of Pt/Pd thermocouples may therefore be more amenable to extrapolation than that of Pt/Pt-Rh thermocouples. Here, the melting temperatures of a Co?CC and a Pd?CC eutectic fixed point are determined by extrapolating the deviation functions of several Pt/Pd thermocouples, after the fashion of Edler et al. The results are compared with the melting temperatures measured using non-contact radiation thermometry. The expanded uncertainty (k = 2) of the melting temperatures determined by extrapolation of the Pt/Pd thermocouple calibrations is ±0.32 °C for the Co?CC fixed point, and ±0.49 °C for the Pd?CC fixed point. For both fixed points, these uncertainties are comparable to those of non-contact radiation thermometry measurements. While a number of assumptions are made in performing the extrapolation of the calibrations, the method does appear to offer a useful complement to non-contact radiation thermometry measurements.     

Temperature dependence of the effective permeability of heat treated Sendust alloys

The temperature dependence (-10°C-70°C) of the effective permeability (μeff) of Sendust alloys (Fe-9-10 wt% Si-5-7 wt% Al) quenched at a room temperature from the various temperatures (400°C-700°C) was investigated at every 2°C in the temperature range of -10°C to 70°C. Most of the studied alloys gave a peak of μeffin the studied temperature range. The peak temperature (Tp) giving the peak μeffvaried with different alloy composition. The alloys of Tp = 20°C are supposed to correspond to the alloys of the zero magnetocrystalline anisotropy constant (K1= 0) at 20°C. The deduced K1= 0 lines at various temperatures (-10°C-50°C) were obtained for an Fe-Si-Al ternary system. The values of peak μeffat the same Tp were different, depending on alloy composition, This difference is due to the difference in the polycrystalline magnetostriction constant (λ s) of the alloys. The peak temperatures of the studied alloys vary with the different heat treatments. This variation is due to the variation of K1, induced by the change in microstructures of the alloys. The variation of the values of peak μeffwith heat treatments was small compared to the difference of those in the alloys of different composition. It seems that the variation of the λ s with heat treatments would be small. Both the compositional dependence of μeffat 20°C and the temperature sensitivity (Δμeff/ΔT) were obtained for both the alloys cooled continuously to room temperature and those quenched at room temperature from 400°C.     

Impact of various solid carriers and spray drying on pre/post compression properties of solid SNEDDS loaded with glimepiride: in vitro-ex vivo evaluation and cytotoxicity assessment

Development of self-nanoemulsifying drug delivery systems (SNEDDS) of glimepiride is reported with the aim to achieve its oral delivery. Lauroglycol FCC, Tween-80, and ethanol were used as oil, surfactant, and co-surfactant, respectively as independent variables. The optimized composition of SNEDDS formulation (F1) was 10% v/v Lauroglycol FCC, 45% v/v Tween 80, 45% v/v ethanol, and 0.005% w/v glimepiride. Further, the optimized liquid SNEDDS were solidified through spray drying using various hydrophilic and hydrophobic carriers. Among the various carriers, Aerosil 200 was found to provide desirable flow, compression, dissolution, and diffusion. Both, liquid and solid-SNEDDS have shown release of more than 90% within 10?min. Results of permeation studies performed on Caco-2 cell showed that optimized SNEDDS exhibited 1.54 times higher drug permeation amount and 0.57 times lower drug excretion amount than that of market tablets at 4?hours (p?p?>?.05, i.e. 0.74). The formulation was found stable with temperature variation and freeze thaw cycles in terms of droplet size, zeta potential, drug precipitation and phase separation. Crystalline glimepiride was observed in amorphous state in solid SNEDDS when characterized through DSC, PXRD, and FT-IR studies. The study revealed successful formulation of SNEDDS for glimepiride.