Biomedical subjects

T Osterberg

Publications and source records attributed to T Osterberg.

At least 19 recordsLinked to original sources

PubMed2005

In vitro models for the blood-brain barrier.

The aim of the present study was to identify a model for the blood-brain barrier based on the use of a continuous cell line, and to investigate the specificity of this model. A set of test compounds, reflecting different transport mechanisms and different degrees of permeability, as well as different physiochemical properties was selected. In vivo data for transport across the blood-brain barrier of this set of test compounds was generated as part of the study using two different in vivo models. A computational prediction model was also developed, based on 74 proprietary Pharmacia compounds, previously tested in one of the in vivo models. Molsurf descriptors were calculated and 21 descriptors were correlated with log(Brain(conc.)/Plasma(conc.)) using partial least squares projection to latent structures (PLS). However, the correlation between predicted and measured values was found to be rather low and differed between one and two log units for several of the compounds. The test compounds were analyzed in vitro using primary bovine and human brain endothelial cells co-cultured with astrocytes, and also using two different immortalized brain endothelial cell lines, one originating from rat and one from mouse. Cell models using cells not derived from the blood-brain barrier, ECV/C6, MDCK and Caco-2 cell lines, were also used. No linear correlation between in vivo and in vitro permeability was found for any of the in vitro models when all compounds were included in the analysis. The highest r2 values were seen in the bovine brain endothelial cells (r2=0.43) and MDCKwt (r2=0.46) cell models. Higher correlations were seen when only passively transported compounds were included in the analysis, bovine brain endothelial cells (r2=0.74), MDCKwt (r2=0.65) and Caco-2 (r2=0.86). By plotting in vivo Papp values against logDpH7.4 it was possible to classify compounds into four different classes: (1) compounds crossing the blood-brain barrier by passive diffusion, (2) compounds crossing the blood-brain barrier by blood-flow limited passive diffusion, (3) compounds crossing the blood-brain barrier by carrier mediated influx, and (4) compounds being actively excreted from the brain by active efflux. Papp and Pe values obtained using the different in vitro models were also plotted against logDpH7.4 and compared to the plot obtained when in vivo Papp values were used. Several of the in vitro models could distinguish between passively distributed compounds and efflux substrates. Of the cell lines included in the present study, the MDCKmdr-1 cell line gave the best separation of passively and effluxed compounds. Ratios between AUC in brain and AUC in blood were also calculated for six of the compounds and compared to ratios between Pe or Papp for transport in the apical to basolateral and basolateral to apical direction. Again the MDCKmdr-1 cell line gave the best correlation with only one compound (AZT) giving large discrepancy between in vitro and in vivo data. None of the in vitro models could identify compounds known to be substrates for carrier mediated influxed as such, and the results indicate that a tighter in vitro blood-brain barrier model probably is needed in order to facilitate studies on carrier mediated influx. The findings presented also indicate that identification of "batteries" of in vitro tests are likely to be necessary in order to improve in vitro-in vivo correlations and to make it possible to perform acceptable predictions of in vivo brain distributions from in vitro data.

Animals↗

PubMed2002

Masticatory ability in 80-year-old subjects and its relation to intake of energy, nutrients and food items.

OBJECTIVE: The aim of the present study was to analyse the relationship between masticatory ability (self-assessed masticatory ability and bite force) and intake of energy, nutrients and food items in a population sample of elderly subjects. DESIGN AND SUBJECTS: From a population sample of 80-year-old people, 160 individuals (74 men and 86 women) took part in an odontological study. MAIN OUTCOME MEASURES: A dental examination including bite force recording, a questionnaire focusing on self-assessed masticatory ability, and a dietary interview. SETTING: Department of Geriatric Medicine, Göteborg University, Sweden. RESULTS: The dental status among the participants varied much (from edentulous in both jaws--22%--to more than 20 natural teeth--30%). The mean maximum bite force was higher in men (165 N) than in women (105 N). Bite force was significantly correlated to the Eichner index and to the number of teeth. One third of the subjects reported no masticatory problem, whereas 18% identified > or = 3 such problems. The intake of energy and nutrients varied much but the means were well above recommended values. The correlations between dental status and bite force on one side and dietary intake on the other side were in general weak and most often statistically non-significant. Impaired general health and reduced dentition were both associated with more masticatory problems. CONCLUSION: The examined sample of 80-year-old subjects had a great variation in dental status, bite force and self-assessed masticatory ability, but these factors had only a minor influence on dietary selection and intake, which on average were well above recommended values.

Aged↗

PubMed2001

Theoretical calculation and prediction of drug transport processes using simple parameters and partial least squares projections to latent structures (PLS) statistics. The use of electrotopological state indices.

A method of modeling and predicting drug transport processes using simple, theoretically computed molecular descriptors and multivariate statistics has been investigated in four data sets related to Caco-2 cell permeability, human intestinal absorption, brain-blood partitioning, and immobilized artificial membrane (IAM) chromatography. The program Molconn-Z was used to compute theoretical molecular descriptors related to electrotopological state indices. Additional parameters related to size and lipophilicity [i.e., calculated molar refraction (CMR) and octanol-water partition coefficient (CLOGP)] were also used in the statistical modeling. Good statistical models were derived (r(2) and Q(2) values ranged from 0.75 to 0.93 and 0.70 to 0.89, respectively) that permit fast computational (electronic) screening and prioritization of virtual libraries.

Blood-Brain Barrier↗

PubMed2001

B-domain deleted recombinant factor VIII formulation and stability.

B-domain deleted recombinant factor VIII (BDDrFVIII) is a deletion form of human coagulation factor VIII. A lyophilized formulation of highly purified BDDrFVIII has been developed that does not require the use of blood-derived products such as human serum albumin (HSA). By avoiding the use of blood-derived products, the BDDrFVIII formulation minimizes the risk of transmitting blood-borne pathogens that may be present in plasma-derived factor VIII or in other recombinant factor VIII products that contain HSA in their formulation. Upon reconstitution with saline (4 mL), the composition of the reconstituted product (62.5 to 250 IU/mL BDDrFVIII) is 18 mg/mL sodium chloride, 3.0 mg/mL sucrose, 1.5 mg/mL L-histidine, 0.25 mg/mL calcium chloride dihydrate, and 0.1 mg/mL polysorbate 80. The optimal combination of these excipients in the lyophilized BDDrFVIII formulation provides long-term stability, as measured by a variety of analytical methods. The formulation preserves factor VIII activity of lyophilized BDDrFVIII during storage for at least 24 months at 8 degrees C, and for up to 6 months at room temperature (25 degrees C). The reconstituted product retains its factor VIII potency for at least 100 hours at 25 degrees C, which would allow it to be continuously administered via an infusion pump, assuming the product is handled under aseptic conditions.

Consumer Product Safety↗

PubMed2001

Chromatographic retention of drug molecules on immobilised liposomes prepared from egg phospholipids and from chemically pure phospholipids.

The partitioning of a chemically diverse set of drugs into liposomes was studied by immobilised liposome chromatography (ILC). For this purpose liposomes composed of (i) purified egg phospholipids (EPL), (ii) synthetic phosphatidylcholine (PC), (iii) PC--synthetic phosphatidylethanolamine (PE) 80:20 (mol/mol) and (iv) PC--synthetic phosphatidylserine (PS) 80:20 (mol/mol) were immobilised in gel beads by freeze-thawing. The drug partitioning was assessed from the retention volume, which was expressed as a capacity factor, K(s), normalised with respect to the amount of immobilised phospholipid. The drug retention on EPL, PC and PC--PE liposomes was very similar, whereas the negatively charged PC--PS liposomes increased the retention of positively charged and decreased retention of negatively charged drugs. The partitioning of drugs on liposome columns (log K(s)) versus their octanol--water partitioning (log P(oct)) showed three separate rectilinear relationships, depending on the charge of the compound (neutral, positive, or negative). Statistical analysis (ANCOVA) proved that the lines had similar slopes. Repeated analysis of four reference compounds showed a low variation (<0.12 log units) over time (about 250 days). A close relationship was observed between the drug retention in short EPL columns with a low content of phospholipids and the retention in longer standard EPL columns. The short 'quick screen bilayer columns' permit analysis of highly lipophilic compounds within 30 min and are thus applicable for medium-throughput screening in drug discovery settings. A very strong rectilinear relationship (r(2)=0.95, n=13) between log K(s) (EPL) and published liposome partitioning data (log D(mem)) confirmed that the ILC drug retention reflects the drug partitioning into the lipid bilayers. A moderate to fair rectilinear relationship was observed between the normalised retention on PC, PC-PE and EPL liposomes (r(2)=0.79, 0.86 and 0.85, respectively, n=24) and corresponding published log k'(IAM) data obtained on immobilised artificial membrane (IAM) columns. Transport across Caco-2 cell monolayers (log P(c)) showed curvilinear relationships with log K(s), log k'(IAM), log P(oct) and log D(oct). The drug fraction absorbed in humans showed a similar relationship to log K(s) values as to surface plasmon resonance signals representing drug-liposome interaction (Danelian et al., 2000 J Med Chem, 43, 2083--2086).

Adrenergic beta-Antagonists↗

PubMed2001

Prediction of drug transport processes using simple parameters and PLS statistics. The use of ACD/logP and ACD/ChemSketch descriptors.

A method of modelling and predicting biopharmaceutical properties using simple theoretically computed molecular descriptors and multivariate statistics has been investigated for several data sets related to solubility, IAM chromatography, permeability across Caco-2 cell monolayers, human intestinal perfusion, brain-blood partitioning, and P-glycoprotein ATPase activity. The molecular descriptors (e.g. molar refractivity, molar volume, index of refraction, surface tension and density) and logP were computed with ACD/ChemSketch and ACD/logP, respectively. Good statistical models were derived that permit simple computational prediction of biopharmaceutical properties. All final models derived had R(2) values ranging from 0.73 to 0.95 and Q(2) values ranging from 0.69 to 0.86. The RMSEP values for the external test sets ranged from 0.24 to 0.85 (log scale).

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed2001

In vitro intestinal permeability of factor Xa inhibitors: influence of chemical structure on passive transport and susceptibility to efflux.

PURPOSE: To study the in vitro intestinal permeability of a number of newly synthesised factor Xa inhibitors to better understand the poor oral absorption of these compounds. METHODS: The bidirectional transport of the fXa inhibitors was studied in the Caco-2 cell model and isolated rat ileal tissue. An attempt was made to characterize efflux mechanisms with the help of commonly used substrates and inhibitors of various transport proteins. In addition, the transport of the fXa inhibitors was studied in MDCK cells transfected with the human MDR1 gene and expressing large amounts of P-glycoprotein (Pgp). RESULTS: The in vitro absorptive permeability was low for all but one of the fXa inhibitors. For compounds with non-substituted amidine, a charge (due to ionisation at neutral pH) may have resulted in poor membrane partitioning. Neutral compounds with substituted amidines were effluxed from the epithelial cells. The significance of the secretion process was illustrated by the results obtained for a neutral analogue showing high absorptive Caco-2 cell permeability that was not obviated by efflux. Transport inhibition studies in Caco-2 and permeability studies in the MDR1-transfected MDCK cells consistently showed that Pgp is not involved in the secretion of fXa inhibitors. Besides efflux, metabolic liability limited the permeation of the neutral lipophilic analogues with a carbamate ester. CONCLUSIONS: Poor intestinal permeability may be an important factor in the incomplete oral absorption of the bisbenzimidazole-type fXa inhibitors. Poor permeability may be related to poor membrane partitioning for hydrophilic analogues, whereas susceptibility to efflux transports and gastro-intestinal enzymatic degradation may limit the permeability of some of the neutral less hydrophilic derivatives.

Animals↗

PubMed2000

Theoretical calculation and prediction of P-glycoprotein-interacting drugs using MolSurf parametrization and PLS statistics.

A method for the modelling and prediction of P-glycoprotein-associated ATPase activity using theoretically computed molecular descriptors and multivariate statistics has been investigated using 22 diverse drug-like compounds. The program MolSurf was used to compute theoretical molecular descriptors related to physicochemical properties such as lipophilicity, polarity, polarizability and hydrogen bonding. The multivariate partial least squares projections to latent structures (PLS) method was used to delineate the relationship between the P-glycoprotein-associated ATPase activity and the theoretically computed molecular descriptors. The PLS analysis of the entire data set, with the exclusion of tamoxifen, resulted in one significant PLS component according to cross-validation with R(2)=0.718, Q(2)=0. 695, S.D.=0.475, F=48.37, RMSE(tr)=0.452, p<0.001. Properties associated with the size of the molecular surface, polarizability and hydrogen bonding had the largest impact on the P-glycoprotein-associated ATPase activity. All these properties should be high to promote high ATPase activity.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed2000

Trends and prognoses of dental status in the Swedish population: analysis based on interviews in 1975 to 1997 by Statistics Sweden.

The aims of this study were to describe changes in dental status over the 22-year period from 1975 to 1997, and to make a prognosis of dental status based on these data for the years 2005 and 2015. The study is based on regular investigations of the living conditions performed by Statistics Sweden of samples varying between 11,582 and 14,964 participants and a response rate from 78% to 86%. The questions of the interview used in this study were focused on dental status and utilization of dental services. The prevalence of edentulism in the age group 25-74 years decreased from 19% in 1975 to 3% in 1996/97. The proportion of dentate persons increased from 75% in 1975 to 97% in 1996/97 in age group 45-64 years with similar trends in the other age groups. In 1996/97, 2.1% of the whole sample (16-84 years) reported that they had received implant-supported restorations. The rate was higher among the elderly and the edentulous subjects. The great regional differences in dental status found in the first part of the observation period remained only in the oldest age group in 1996/97. The prognosis predicts that 95% of the subjects in age group 65-74 years and 90% in age group 75-84 years will be dentate in the year 2015. The substantial increase of dentate subjects among the elderly that has occurred during the past few decades and its expected continuation in the coming years implies a great change in need and demand for dental care services.

Adolescent↗

PubMed1999

Theoretical calculation and prediction of intestinal absorption of drugs in humans using MolSurf parametrization and PLS statistics.

A method for modeling and prediction of the intestinal absorption of drugs in humans using theoretically computed molecular descriptors and multivariate statistics has been investigated using 20 diverse drug-like compounds. The program MolSurf was used to compute theoretical molecular descriptors related to physicochemical properties such as lipophilicity, polarity, polarizability and hydrogen bonding. The multivariate Partial Least Squares Projections to Latent Structures (PLS) method was used to delineate the relationship between the intestinal absorption of drugs in humans and the theoretically computed molecular descriptors.Good statistical models were derived. Properties associated with hydrogen bonding had the largest impact on absorption and should be kept to a minimum to promote high absorption. High charge-transfer properties and the presence of surface electrons, i.e. valence electrons, which are not tightly bonded to the molecule, were also found to promote high absorption.

Computer Simulation↗

PubMed1999

Physical state of L-histidine after freeze-drying and long-term storage.

Liquid samples of L-histidine of varying pH values and mixed with salt, metal ions, polysorbate 80 and sucrose have been analysed by differential scanning calorimetry to evaluate the influence of these additives on the glass transition temperature and crystallisation of L-histidine during freezing and thawing. L-Histidine solutions of varying pH were freeze-dried with and without a thermal cycle and the physical state of the freeze-dried cakes, following long-term storage, were studied by powder X-ray diffraction. Amorphous L-histidine crystallised when it was exposed to moisture, and the identity of the crystalline materials is reported. The crystallisation of L-histidine during freezing and thawing is dependent on the pH of the solution and is shown to be at a minimum at pH 6, which coincides with the pK(a) of the imidazoline function. Sucrose inhibited the crystallisation of L-histidine during thawing, while sodium chloride or polysorbate 80 did not. The addition of metal ions (Ca2+ and Mg2+) up to 10% (w/w) did not depress the glass transition temperature significantly, while the addition of Zn2+ increased it. The physical state of L-histidine after freeze-drying is shown to be dependent on both the pH of the solution and the freezing cycle. The risk of crystallisation of amorphous L-histidine is low if the freeze-dried material is protected from moisture.

Calcium↗

PubMed1998

Theoretical calculation and prediction of brain-blood partitioning of organic solutes using MolSurf parametrization and PLS statistics.

Sixty-three structurally diverse compounds were investigated to statistically model the brain-blood partitioning of organic solutes using theoretically computed molecular descriptors and multivariate statistics. The program MolSurf was used to compute theoretical molecular descriptors related to physicochemical properties such as lipophilicity, polarity, polarizability, and hydrogen bonding. The multivariate Partial Least Squares Projections to Latent Structures (PLS) method was used to delineate the relationship between the brain-blood partitioning of organic solutes and the theoretically computed molecular descriptors. Good statistical models were derived. Properties associated with polarity and Lewis base strength had the largest impact on the blood-brain partitioning and should be kept to a minimum to promote high partitioning. The absence of atoms capable of hydrogen bonding interactions as well as high lipophilicity and the presence of polarizable surface electrons, i.e., valence electrons, were also found to promote high brain-blood partitioning. The results indicate that theoretically computed molecular MolSurf descriptors in conjunction with multivariate statistics of PLS type can be used to successfully model the brain-blood partitioning of organic solutes and hence differentiate drugs with poor partitioning from those with acceptable partitioning at an early stage of the preclinical drug-discovery process.

Blood-Brain Barrier↗

PubMed1998

Self-assessed masticatory ability in relation to maximal bite force and dental state in 80-year-old subjects.

By means of a questionnaire, clinical examination and force recordings, the relationships between self-assessed masticatory ability, dental state and bite force were studied in 160 80-year-old persons, 74 men and 86 women. The subjects were in general satisfied with their masticatory ability and 70% had no problems, while 6% reported three or more problems with mastication. Half of the subjects were dentate without removable prostheses and almost one-third had 20 or more natural teeth. The edentulous persons (about one-fifth of all) reported more problems related to mastication than the other dentition groups. The maximal bite force varied much and exhibited a significant correlation to the number of remaining teeth and dental state. The self-assessed masticatory ability was only weakly correlated with dental state and bite force. It was concluded that many subjects with few or no remaining teeth and/or removable dentures had only few complaints of impaired masticatory function and showed a good adaptation to an impaired dental status and small maximal bite force.

Adaptation, Physiological↗

PubMed1998

Utilization of dental services in relation to socioeconomic and health factors in the middle-aged and elderly Swedish population.

The aims of this study were to describe the change in reported time since the latest visit to a dentist between the years 1980/81 and 1988/89 and the reported use of dental services in relation to age, dental state, and socioeconomic and health characteristics in a sample of the Swedish population in 1988/89. The studies are based on interviews by Statistics Sweden about the living conditions. In the investigations in 1980/81, 14,964 inhabitants between 16 and 84 years of age participated, and in 1988/89, 13,309 inhabitants. In all age groups there was a significantly higher frequency of reported visits to a dentist last year in 1988/89 than in 1980/81. In the age group 50-64 years old this figure increased from 54% to 75%, and in the age group 65-84 years old it increased from 26% to 39%. In the investigation in 1988/89 about 75% of the dentulous women in all age groups up to 75 years reported visiting a dentist last year. The relative risk for not visiting a dentist last year, adjusted for age, gender, and dental state, was higher in dentulous subjects with low income and education, not married, not native-born, living in rural areas, smoking, and low social and physical activity. The results of the logistic regression analysis showed that, among the elderly, functional ability and general health factors have lower significance for time since last visit to a dentist than socioeconomic, social support, and life-style factors.

Activities of Daily Living↗

PubMed1998

Root caries and some related factors in 88-year-old carriers and non-carriers of Streptococcus sobrinus in saliva.

The prevalence of caries on exposed root surfaces in 88-year-old subjects with and without salivary levels of Streptococcus sobrinus was studied. Ninety-two individuals were examined with regard to root caries lesions and fillings. The root caries index (RCI) was calculated and related to salivary flow rate and buffer capacity, plaque score and salivary counts of Streptococcus mutans, S. sobrinus and lactobacilli. In 89 subjects with exposed root surfaces, all but 2 harbored mutans streptococci; 51 subjects carried S. mutans only, 35 both S. sobrinus and S. mutans, and 1 S. sobrinus only. The RCI was significantly higher in persons with than those without S. sobrinus (p < 0.05). Subjects with both S. sobrinus and S. mutans had higher counts of total mutans streptococci and lactobacilli than subjects with only S. mutans (p < 0.05). The RCI was significantly correlated to S. sobrinus and S. mutans (p < 0.05). The positive correlation between the RCI and S. sobrinus was still significant when the other tested variables were kept constant, whereas the correlation between the RCI and S. mutans was weaker when S. sobrinus and lactobacilli were kept constant. The D-component of the RCI (DSr%) was significantly correlated to S. sobrinus, S. mutans and lactobacilli, whereas the F-component of the RCI showed no significant correlation to any of the tested variables. A stepwise multiple correlation showed that the variance of DSr% was best explained in the S. sobrinus carriers by S. sobrinus and the salivary buffer effect, and in the non-carriers by S. mutans.

Aged↗

PubMed1997

Recombinant factor VIII SQ--inactivation kinetics in aqueous solution and the influence of disaccharides and sugar alcohols.

PURPOSE: To investigate the influence of various nonreducing disaccharides and sugar alcohols on the inactivation kinetics of recombinant factor VIII SQ (r-VIII SQ) in aqueous solution not containing albumin as a stabiliser. METHODS: The stability of r-VIII SQ was followed using measurement of activity (VIII:C) and HPLC gel filtration at different temperatures. The thermal stability was investigated using differential scanning calorimetry (DSC). RESULTS: The decline in VIII:C followed pseudo-first order kinetics. However, the Arrhenius plot was not linear for formulations without carbohydrate, demonstrating a distinct, reproducible curvature. The reaction rate at 5 degrees C was faster than expected from the Arrhenius kinetics. The energy of activation (Ea) for formulations without added carbohydrates, derived from the linear part of the Arrhenius plot, varied between 77 and 86 kJ/mole in the temperature range 20-37 degrees C. The addition of 600 mg/ml sucrose increased the Ea to 104 kJ/mole. DSC measurements showed that Tm' was 64.2 +/- 0.2 degrees C for r-VIII SQ without stabiliser. This value increased linearly with increasing concentrations of carbohydrate. This stabilising effect is most probably explained by the theory of preferential hydration. CONCLUSIONS: The inactivation kinetics of r-VIII SQ in aqueous solution without addition of carbohydrates followed pseudo-first order kinetics but the Arrhenius plot was nonlinear. Sucrose and sorbitol both had highly stabilising effects on r-VIII SQ at concentrations above 300 mg/ml. The preparation containing 600 mg/ml sucrose was stable for at least 12 months at 5 degrees C and 6 months at 25 degrees C.

Animals↗

PubMed1997

Theoretical calculation and prediction of Caco-2 cell permeability using MolSurf parametrization and PLS statistics.

PURPOSE: To statistically model the permeability across Caco-2 cell monolayers using theoretically computed molecular descriptors and multivariate statistics. METHODS: Seventeen structurally diverse compounds were investigated. The program MolSurf was used to compute theoretical molecular descriptors related to physico-chemical properties such as lipophilicity, polarity, polarizability and hydrogen bonding. The multivariate Partial Least Squares Projections to Latent Structures (PLS) method was used to delineate the relationship between the permeability across Caco-2 cell monolayers and the theoretically computed molecular descriptors. RESULTS: Excellent statistical models were derived. Properties associated with hydrogen bonding had the largest impact on diffusion through the monolayers and should be kept at a minimum to promote high permeability. High lipophilicity and the presence of surface electrons, i.e. valence electrons, which are not tightly bonded to the molecule, were also found to have a favorable influence to achieve high permeability. CONCLUSIONS: The results indicate that theoretically computed molecular MolSurf descriptors in conjunction with multivariate statistics of PLS type can be used to successfully model permeability across Caco-2 cell monolayers and, thus, differentiate drugs with poor permeability from those with acceptable permeability at an early stage of the preclinical drug discovery process.

Caco-2 Cells↗

PubMed1997

Development of freeze-dried albumin-free formulation of recombinant factor VIII SQ.

PURPOSE: To develop a stable freeze-dried formulation of recombinant factor VIII-SQ (r-VIII SQ) without the addition of albumin. METHODS: Different formulations were evaluated for their protective effect during sterile filtration, freeze-thawing, freeze-drying, reconstitution and long term storage. Factor VIII activity (VIII:C), visual inspection, clarity, solubility, moisture content and soluble aggregates and/ or fragments were assayed. RESULTS: A combination of non-crystallising excipients (L-histidine and sucrose), a non-ionic surfactant (polysorbate 80) and a crystalline bulking agent (sodium chloride) was found to preserve the factor VIII activity during formulation, freeze-drying and storage. Calcium chloride was included to prevent dissociation of the heavy and light chains of r-VIII SQ. Sodium chloride was chosen as the primary bulking agent since the concentration of sodium chloride necessary for dissolution of r-VIII SQ in the buffer will inhibit the crystallization of many potential cake formers. It was found that L-histidine, besides functioning as a buffer, also protected r-VIII SQ during freeze-drying and storage. A pH close to 7 was found to be optimal. Some potential macromolecular stabilisers, PEG 4000, Haes-steril and Haemaccel, were evaluated but they did not improve the recovery of VIII:C. The freeze-dried formulation was stable for at least two years at 7 degrees C and for at least one year at 25 degrees C. The reconstituted solution was stable for at least 100 hours at 25 degrees C. CONCLUSIONS: The albumin-free formulation resulted in consistently high recovery of VIII:C, very low aggregate formation and good storage stability. The stability of the reconstituted solution makes the formulation suitable for continuous administration via infusion pump. The formulation strategy described here may also be useful for other proteins which require a high ionic strength.

Adsorption↗