Lawrence H. Block

Lyophilization of Cationic Lipid–protamine–DNA (LPD) Complexes

Cationic lipid-based gene delivery systems have shown promise in transfecting cells both in vitro and in vivo. However, these systems tend to form aggregates in liquid formulation during storage, which has limited their clinical applications. As a result, lyophilization of these systems has recently become a subject of increasing interest. In this paper, lyophilization of LPD, a novel cationic lipid-based gene delivery system, was studied. Both particle size and transfection efficiency could be preserved in the presence of sufficient amount of appropriate lyoprotectant. A series of monosaccharides and disaccharides, including dextrose, galactose, mannose, lactose, maltose, sucrose and trehalose, were evaluated for their lyoprotective effect and disaccharides showed more superior protection to monosaccharides. The effect of different freezing protocols for lyophilization was also evaluated and no significant difference was found. However, for freeze-thawing, fast freezing caused less aggregation. Finally, nonlyophilized LPD and LPD lyophilized with 10% sucrose were stored at different temperatures and their stability was followed for eight weeks. Lyophilized LPD could be stored at room temperature without significant change in particle size or loss of transfection efficiency.

Chitosan processing: influence of process parameters during acidic and alkaline hydrolysis and effect of the processing sequence on the resultant chitosan’s properties

Abstract The objectives of this study were to investigate the influence of reaction temperature, processing time, and mechanical shear on the depolymerization (DP) of chitosans and to evaluate the importance of the sequencing of the deacetylation and DP processes on the resultant macromolecules and their properties. Process sequence did not alter the degree of deacetylation (DD), the intrinsic viscosity, or the molecular weight (MW). Treatment conditions affected the properties of the resultant polymer: the reaction temperature and processing time had a significant impact on the MWs and DDs of the resultant polymer. Mechanical shear, however, did not significantly affect the above properties. Furthermore, polymer crystallinity was affected by reaction temperature but not by shear or processing time.

The use of chitosan gels as matrices for electrically-modulated drug delivery.

This study evaluated and characterized the use of chitosan gels as matrices for electrically modulated drug delivery. Chitosan gels were prepared by acetylation of chitosan and subsequently hydrated to facilitate further studies. After determining the degree of deacetylation, hydrated and unhydrated gel formulations were characterized for their microviscosity and compression strength. In the electrification studies, gel mass variation, surface pH changes, and later, release-time profiles for neutral (hydrocortisone), anionic (benzoic acid), and cationic (lidocaine hydrochloride) drug molecules from hydrated chitosan gels were monitored in response to different milliamperages of current as a function of time. Hydrated gels had very similar microviscosity while exhibiting differences in the gel strength, results which are not inconsistent as they pertain to different aspects of the gel. The cumulative gel mass loss and rate of gel mass loss increased with an increase in the milliamperage (mA) of the applied current. Gel syneresis - principally involving electroosmosis and gel collapse - was pronounced, particularly at higher mAs and for chitosan gels with lower degrees of acetylation. The surface pH values of the gels were lower at the anode and higher at the cathode, in accordance with reports in the literature. The release of the model drugs from the gel matrix was in the order benzoic acid>hydrocortisone>lidocaine, which is consistent with the electrokinetically competing forces that are involved in these gels. Adequate characterization of electrical effects on formulation matrices, such as chitosan gels, is critical to the development of effective and reliable electrically modulated drug delivery systems.

Chitosan as an enabling excipient for drug delivery systems: I. Molecular modifications

Chitosan was physicochemically modified for its potential use as a matrix for an implantable antibiotic delivery system that could sustain bactericidal concentrations in the vicinity of an implant or prosthesis. Deacetylation and depolymerization of chitosan were implemented in order to increase the number or accessibility of the reactive amino groups on the polymer backbone for better polymer-drug interaction. The deacetylation process involved reaction of particulate chitosan/depolymerized chitosan with alkali. The rate of deacetylation of chitosan was directly proportional to the reaction temperature up to 80 degrees C; beyond 80 degrees C, rapid degradation of the polymer occurred. The depolymerization of chitosan involved acid digestion of the polymer followed by application of mechanical agitation. This depolymerized product, although water insoluble, possessed a molecular weight that was one to two orders of magnitude lower than that of commercially available chitosans. These products not only exhibited improved reactivity, but also showed increased crystallinity when compared with the parent chitosan. The reactivity was found to be inversely proportional to chitosans molecular weight. The depolymerization and deacetylation treatments afforded formation of chitosan having a greater number of amino groups available for interactions with the anionic actives.

Use of Chitosan in Compressed Tablets of Diclofenac Sodium: Inhibition of Drug Release in an Acidic Environment

The purpose of this study was to evaluate the potential utility of chitosan (I) in inhibiting diclofenac sodium (II) release in the gastric environment from a directly compressible tablet formulation. I, subjected to depolymerization to improve its microcrystallinity and subsequent compressibility, was then used to prepare tablets of II. A full-factorial design was employed to evaluate the effects of degree of N-deacetylation of I, and the pH and ionic strengths, mu, of the dissolution media on drug release. Directly compressible tablets were prepared from admixtures of 25 mg of II, 174 mg of I of various degrees of N-deacetylation (74, 87, and 92%), and 1 mg of magnesium stearate. The in vitro dissolution studies were performed using aqueous buffers (pHs 1.2, 3.8, and 6.8, and mu of approximately 1.0 and 0.1). The slopes of logarithmically transformed cumulative percent released-time curves (from t = 0 to t = 5 hr) were compared. Analyses of variance performed using SAS indicated that the degree of N-deacetylation of chitosan significantly affected drug release at pHs 1.2 and 6.8 (p < 0.0001). An increase in the pH of the dissolution medium resulted in an increase in drug release (p < 0.0001). The ionic strength of the dissolution medium did not significantly affect drug release at any of the pHs studied (p > 0.198). Besides the poor aqueous solubility of II, the two factors possibly affecting the drug release in the acidic environment were (a) the formation of a rate-limiting chitosan gel barrier; and (b) the ionic interaction of II with ionized amino groups of I.

Evaluation of Ethylcellulose as A Matrix for Prolonged Release Formulations. I. Water Soluble Drugs: Acetaminophen and Theophylline

AbstractIn this study ethylcellulose was evaluated as a carrier for the preparation of prolonged release solid dispersions of relatively water soluble drugs, acetaminophen and theophylline. The solid dispersions containing various concentrations (7.5, 15.0 and 30.0 % by weight of drug) of ethylcellulose of different viscosity grades (21, 95, 209 and 350 cps) were prepared by the solvent method. The concentration of polymer in the formulation was the determining factor in controlling release rate of the drug, as the results indicate prolongation in release of the drug with increase in amount of ethylcellulose. The higher the viscosity grade of ethylcellulose, slower the release of drug from the solid dispersions. The release of drug from the tablets was more prolonged compared to the granular solid dispersions. In vitro release of acetaminophen and theophylline was more or less similar in both dissolution media. The viscosity grade of ethylcellulose showed slight influence on the release rate of drug from ...

Spray-dried chitinosans. Part I: preparation and characterization.

PURPOSE Physicochemical and micromeritic characterization of chitinosans. METHODS Chitinosans subjected to N-deacetylation and depolymerization were characterized for degree of N-deacetylation (DD), molecular weight (MW), pK(a), particle size determination and morphology, tap/bulk density measurements, surface area determinations, and determination of flow properties. RESULTS The chitinosan DDs and MWs were dependent on the processing conditions and ranged from 66 to 89% and 2-522 kDa, respectively. Chitinosan particle sizes and shapes were dependent on drying conditions (range 8-465 microm). Spray-dried chitinosans were spherical and had smaller particle sizes than the non-spray-dried materials which were irregularly shaped particles. Higher density values were obtained for processed materials than those for the raw material. Lower specific surface areas were observed for non-spray-dried chitinosans (0.28-1.59 m(2)/g) than for spray-dried chitinosans (0.74-3.01 m(2)/g). Weight variation of chitinosan tablets indicated that spray-dried chitinosans possessed improved flow characteristics as compared with tray-dried chitinosans. CONCLUSIONS The effect of drying method employed in chitinosan manufacture, i.e. spray versus tray drying, on the physicochemical and micromeritic properties of the resultant chitinosans were evaluated. Although the drying methods did not significantly influence the physicochemical properties, they affected the micromeritic properties of the resultant chitinosans.

Spray-dried chitinosans. Part II: in vitro drug release from tablets made from spray-dried chitinosans.

PURPOSE Application of spray-dried chitinosans as excipients for use in drug delivery systems was explored. METHODS Spray- and tray-dried chitinosans previously N-deacetylated and depolymerized were used. Directly compressed tablets (200mg) containing tetracycline, chitinosan, and magnesium stearate were prepared. The tablets were characterized for dimensions, weight, friability, crushing strengths, disintegration, and dissolution. RESULTS The tablet weights, thickness, and diameters were not affected by the chitinosan selected (P>0.05). Friability of tablets containing tray-dried chitinosans was generally higher (and crushing strengths were lower) than tablets containing spray-dried chitinosans. Chitinosan molecular weight, degree of N-deacetylation, and drying method used, significantly affected crushing strengths (P<0.0001). Disintegration times were affected only by the type of chitinosan (P<0.0001) but not by the drying method used (P>0.9). Dissolution from tablets was significantly affected by the chitinosan type (P<0.025), but not affected by the drying method (P>0.5). CONCLUSIONS Spray drying improved binding functionality of chitinosans, thereby enhancing the tablet crushing strength; however, friability, disintegration, and dissolution profiles were not significantly affected. The data obtained from this study support the usefulness of spray-dried chitinosans as excipients for use in drug delivery systems.

Evaluation of Ethylcellulose as a Matrix for Prolonged Release Formulations. II. Sparingly Water-Soluble Drugs: Ibuprofen and Indomethacin

AbstractThe main purpose of this investigation was to evaluate ethylcellulose as a carrier for the preparation of prolonged release solid dispersions of sparingly water-soluble drugs, ibuprofen and indomethacin. Solid dispersions containing various concentrations of ethylcellulose of different viscosity grades were prepared by the solvent method. Tablets were directly compressed from solid dispersions (40/100 mesh) with 0.5% Primojel as a disintegrant and 0.5% magnesium stearate as a lubricant. In vitro release studies employed a rotating bottle system with Sorensons buffer solution (pH 7.4). It was found that prolongation of drug release was primarily associated with an increase in amount of ethylcellulose rather than the viscosity grade. Nonetheless, the higher the viscosity grade of ethylcellulose, the slower the release of drug from granular and compressed solid dispersions. The release rate of ibuprofen was faster than that of indomethacin from different solid dispersion formulations.

Rheological evaluation of inter-grade and inter-batch variability of sodium alginate.

Polymeric excipients are often the least well-characterized components of pharmaceutical formulations. The aim of this study was to facilitate the QbD approach to pharmaceutical manufacturing by evaluating the inter-grade and inter-batch variability of pharmaceutical-grade polymeric excipients. Sodium alginate, a widely used polymeric excipient, was selected for evaluation using appropriate rheological methods and test conditions. The materials used were six different grades of sodium alginate and an additional ten batches of one of the grades. To compare the six grades, steady shear measurements were conducted on solutions at 1%, 2%, and 3% w/w, consistent with their use as thickening agents. Small-amplitude oscillation (SAO) measurements were conducted on sodium alginate solutions at higher concentrations (4–12% w/w) corresponding to their use in controlled-release matrices. In order to compare the ten batches of one grade, steady shear and SAO measurements were performed on their solutions at 2% w/w and 8% w/w, respectively. Results show that the potential interchangeability of these different grades used as thickening agents could be established by comparing the apparent viscosities of their solutions as a function of both alginate concentration and shear conditions. For sodium alginate used in controlled-release formulations, both steady shear behavior of solutions at low concentrations and viscoelastic properties at higher concentrations should be considered. Furthermore, among batches of the same grade, significant differences in rheological properties were observed, especially at higher solution concentrations. In conclusion, inter-grade and inter-batch variability of sodium alginate can be determined using steady shear and small-amplitude oscillation methods.