Effect of Structurally Related Compounds on Desupersaturation Kinetics of Indomethacin.

PURPOSE: The pharmaceutical literature contains examples wherein desupersaturation from high concentrations does not proceed to equilibrium concentration of the thermodynamically most stable form but remains above equilibrium. The purpose of the current research was to investigate the effect of structurally related compounds on desupersaturation kinetics as a possible explanation for a higher than equilibrium solubility after crystal growth of γ-indomethacin (γ-IMC). METHODS: Three structurally related compounds (SRC) - cis-sulindac (c-SUL), trans-sulindac (t-SUL) and indomethacin-related compound-A (IMC-A) -were investigated. Desupersaturation kinetics to the most stable γ-IMC, in the presence of c-SUL, t-SUL or IMC-A, was measured at pH 2.0. RESULTS: The SRCs c-SUL and t-SUL were effective crystallization inhibitors of IMC, while IMC-A was not a potent crystallization inhibitor of IMC. Among the sulindac isomers, t-SUL was a stronger crystallization inhibitor. The apparent solubility of γ-IMC crystals grown from supersaturated solutions in the presence of SRCs matched the equilibrium solubility of γ-IMC. During crystallization of IMC in the presence of IMC-A, the concentration of IMC-A declined initially but rebounded as supersaturation and crystallization rate of IMC declined, suggesting that IMC-A itself became incorporated in the IMC crystal lattice at higher degrees of IMC supersaturation. CONCLUSIONS: The results suggest that high apparent solubility after crystallization of IMC reported by several authors is not related to the presence of IMC-A impurity. The greater IMC crystal growth rate inhibition by t-SUL than by c-SUL was consistent with the proposed orientation of SUL molecules adsorbed on the IMC crystal, providing a mechanistic understanding of the inhibition.

Mechanisms for the Slowing of Desupersaturation of a Weak Acid at Elevated pH.

Supersaturating drug delivery systems are used to achieve higher oral bioavailability for poorly soluble drugs. However, supersaturated solutions often decline to lower concentrations by precipitation and crystallization. The purpose of the current research is to provide a mechanistic understanding of drug crystallization as a function of pH, using indomethacin (IMC, pKa 4.18) as a model compound. Desupersaturation kinetics to the γ-form of IMC was measured at pH 2.0, 3.0, 4.0, and 4.5 from an initial degree of supersaturation of 2.5-6. At equivalent levels of supersaturation, crystal growth rates decreased with an increase in solution pH. Two mechanisms for this phenomenon, reactive diffusion (resulting in a higher surface pH as compared to bulk pH) and inhibition of crystallization by structurally similar ionized IMC at higher pH, were explored. Non-steady-state models for reactive diffusion showed that the surface pH was only 0.01 units above that of the bulk solution pH. Mass transport models for reactive diffusion during crystallization could not explain the decrease in desupersaturation kinetics at higher pH. However, zeta potentials as high as -70 mV suggested that IMC- is adsorbed on the surface of the particles. A mathematical model for inhibition of crystal growth by IMC- accounted for the pH effect suggesting that ionized IMC acts as an effective crystallization inhibitor of IMC.

Evaluation of Predictors of Protein Relative Stability Obtained by Solid-State Hydrogen/Deuterium Exchange Monitored by FTIR.

PURPOSE: Hydrogen/deuterium (H/D) exchange over a range of temperatures suggests a protein structural/mobility transition in the solid state below the system glass transition temperature (Tg). The purpose of this study was to determine whether solid-state protein stability correlates with the difference between storage temperature and apparent Td where an abrupt change in mobility occurs, or alternatively, the extent of H/D exchange at a single temperature correlates directly to protein stability in lyophilized solids. METHODS: Solid-state H/D exchange was monitored by FTIR spectroscopy to study the extent of exchange and the apparent transition temperature in both pure recombinant human serum albumin (rHSA) and rHSA formulated with sucrose or trehalose. H/D exchange of freeze-dried formulations at 11% RH and temperatures from 30 to 80°C was monitored. Protein stability against aggregation at 40°C/11% RH for 6 months was assessed by size exclusion chromatography (SEC). RESULTS: Both sucrose and trehalose showed equivalent protection of protein secondary structure by FTIR. The rHSA:sucrose formulation showed superior long-term stability at 40°C by SEC over the trehalose formulation, but the apparent Td determined from H/D exchange was much higher in the trehalose formulation. Instead, the extent of H/D exchange (X∞) was lower in the sucrose formulation at the temperature of the stability studies (40°C) than found for the trehalose formulation, which was consistent with better stability in the sucrose formulation. CONCLUSIONS: While apparent Td did not correlate with protein stability for rHSA, the extent of H/D exchange, X∞, did.

Oral Ingestion and Intraventricular Injection of Curcumin Attenuates the Effort-Related Effects of the VMAT-2 Inhibitor Tetrabenazine: Implications for Motivational Symptoms of Depression.

Effort-related choice tasks are used for studying depressive motivational symptoms such as anergia/fatigue. These studies investigated the ability of the dietary supplement curcumin to reverse the low-effort bias induced by the monoamine storage blocker tetrabenazine. Tetrabenazine shifted effort-related choice in rats, decreasing high-effort lever pressing but increasing chow intake. The effects of tetrabenazine were reversed by oral ingestion of curcumin (80.0-160.0 mg/kg) and infusions of curcumin into the cerebral ventricles (2.0-8.0 µg). Curcumin attenuates the effort-related effects of tetrabenazine in this model via actions on the brain, suggesting that curcumin may be useful for treating human motivational symptoms.

Spatial Variation of Pressure in the Lyophilization Product Chamber Part 2: Experimental Measurements and Implications for Scale-up and Batch Uniformity.

Product temperature during the primary drying step of freeze-drying is controlled by a set point chamber pressure and shelf temperature. However, recent computational modeling suggests a possible variation in local chamber pressure. The current work presents an experimental verification of the local chamber pressure gradients in a lab-scale freeze-dryer. Pressure differences between the center and the edges of a lab-scale freeze-dryer shelf were measured as a function of sublimation flux and clearance between the sublimation front and the shelf above. A modest 3-mTorr difference in pressure was observed as the sublimation flux was doubled from 0.5 to 1.0 kg·h-1·m-2 at a clearance of 2.6 cm. Further, at a constant sublimation flux of 1.0 kg·h-1·m-2, an 8-fold increase in the pressure drop was observed across the shelf as the clearance was decreased from 4 to 1.6 cm. Scale-up of the pressure variation from lab- to a manufacturing-scale freeze-dryer predicted an increased uniformity in drying rates across the batch for two frequently used pharmaceutical excipients (mannitol and sucrose at 5% w/w). However, at an atypical condition of shelf temperature of +10°C and chamber pressure of 50 mTorr, the product temperature in the center vials was calculated to be a degree higher than the edge vial for a low resistance product, thus reversing the typical edge and center vial behavior. Thus, the effect of local pressure variation is more significant at the manufacturing-scale than at a lab-scale and accounting for the contribution of variations in the local chamber pressures can improve success in scale-up.

Large-Scale Freeze-Thaw of Protein Solutions: Study of the Relative Contributions of Freeze-Concentration and Ice Surface Area on Stability of Lactate Dehydrogenase.

Although bulk biotherapeutics are often frozen during fill finish and shipping to improve their stability, they can undergo degradation leading to losses in biological activity during sub-optimal freeze-thaw (F/T) process. Except for a few small-scale studies, the relative contribution of various F/T stresses to the instability of proteins has not been addressed. Thus, the objective of this study was to determine the individual contributions of freeze-concentration, ice surface area, and processing time to protein destabilization at a practical manufacturing-scale. Lactate dehydrogenase (LDH) in histidine buffer solutions were frozen in 1L containers. The frozen solutions were sliced into representative samples and assessed for the ice specific surface area (SSA) and extent of solutes freeze-concentration. For the first time to our knowledge, ice SSA was measured in dried samples from large-volume protein solutions using volumetric nitrogen adsorption isotherms. SSA measurements of the freeze-dried cakes showed that the ice surface area increased with an increase in the freezing rate. The ice SSA was also impacted by the position of the sample within the container: samples closer to the active cooled surface of the container exhibited smaller ice surface area compared to ice-cored samples from the center of the bottle. The freeze-concentrate composition was determined by measuring LDH concentration in the ice-cored samples. The protein distributed more evenly throughout the frozen solution after fast freezing which also correlated with enhanced protein stability compared to slow freezing conditions. Overall, better protein stability parameters correlated with higher ice SSA and lower freeze-concentration extent which was achieved at a faster freezing rate. Thus, extended residence time of the protein at the freeze-concentrated microenvironment is the critical destabilizing factor during freezing of LDH in bulk histidine buffer system. This study expands the understanding of the relative contributions of freezing stresses which, coupled with the knowledge of cryoprotection mechanisms, is imperative to the development of optimized processes and formulations aiming stable frozen protein solutions.

Impact of controlled ice nucleation and lyoprotectants on nanoparticle stability during Freeze-drying and upon storage.

The freezing step of the lyophilization process can impact nanoparticle stability due to increased particle concentration in the freeze-concentrate. Controlled ice nucleation is a technique to achieve uniform ice crystal formation between vials in the same batch and has attracted increasing attention in pharmaceutical industry. We investigated the impact of controlled ice nucleation on three types of nanoparticles: solid lipid nanoparticles (SLNs), polymeric nanoparticles (PNs), and liposomes. Freezing conditions with different ice nucleation temperatures or freezing rates were employed for freeze-drying all formulations. Both in-process stability and storage stability up to 6 months of all formulations were assessed. Compared with spontaneous ice nucleation, controlled ice nucleation did not cause significant differences in residual moisture and particle size of freeze-dried nanoparticles. The residence time in the freeze-concentrate was a more critical factor influencing the stability of nanoparticles than the ice nucleation temperature. Liposomes freeze-dried with sucrose showed particle size increase during storage regardless of freezing conditions. By replacing sucrose with trehalose, or adding trehalose as a second lyoprotectant, both the physical and chemical stability of freeze-dried liposomes improved. Trehalose was a preferable lyoprotectant than sucrose to better maintain the long-term stability of freeze-dried nanoparticles at room temperature or 40 °C.

Spontaneous crystalline-to-amorphous phase transformation of organic or medicinal compounds in the presence of porous media, part 3: effect of moisture.

PURPOSE: Amorphization of crystalline compounds using mesoporous media is a promising technique to improve the solubility and drug release of poorly-soluble compounds. The objective of this paper is to understand the effect of moisture on the capacity and performance of vapor-phase mediated amorphization. METHODS: Mesoporous silicon dioxide (SiO(2)) and crystalline naphthalene were used as the model system. The effect of moisture on the amorphization capacity of naphthalene was determined using adsorption chambers with various levels of relative humidity. Enthalpy and capacity of water vapor adsorption on SiO(2) were measured using isothermal microcalorimetry and thermogravimetry. RESULTS: Moisture not only suppressed the amorphization capacity of naphthalene, but reversed an already-amorphized formulation as well. On the other hand, through the same competitive interaction, improved drug release and enhanced solubility were obtained. The initial supersaturation was followed by an entropy-driven crystallization. In addition, moisture-induced siloxane bond fracture was found at normal processing conditions, which led to the changes in silica surface chemistry. However, the implication in amorphization has not reached a definitive conclusion. CONCLUSIONS: Humidity during processing and storage must be carefully controlled for this type of amorphous formulation. Further investigation is needed to better understand the moisture-induced changes of silica.

Differential heat of adsorption of water vapor on silicified microcrystalline cellulose (SMCC): an investigation using isothermal microcalorimetry.

A novel dual-shaft configuration in isothermal microcalorimetry was developed to study the interaction of water vapor with pharmaceutical excipients. An instrument performance test is suggested to validate the experimental data. Reliable experimental results can be collected using a single perfusion shaft; however, there was limitation of the dual-shaft configuration, which resulted deviation in the experimental results. A periodic performance test is recommended. Silicified microcrystalline cellulose (SMCC) was used as a model system to study the interaction using the dual-shaft method. Enthalpy of water vapor adsorption on SMCC was determined and compared to literature data. The data collected using the dual-shaft configuration did not reflect the actual physical system. The deviation was most likely due to the lack of flow control caused by viscous resistance. The enthalpy of adsorption was then calculated using isothermal microcalorimetry coupled with a dynamic vapor sorption apparatus. The results, -55 kJ/mol at low relative humidity (RH) to -22 kJ/mol at high RH, were consistent with the physical phenomenon of water vapor adsorption. Enthalpy of adsorption showed surface heterogeneity of SMCC and suggested multilayer condensation of water at approximately 60% RH. However, at high RH, the results showed the moisture-excipient interaction can be more complex than the proposed mechanism.

Aqueous solubility of crystalline and amorphous drugs: Challenges in measurement.

Measurement of drug solubility is one of the key elements of active pharmaceutical ingredient (API) characterization during the drug discovery and development process. This report is a critical review of experimental methods reported in the literature for the measurement of aqueous solubility of amorphous, partially crystalline and crystalline organic compounds. A summary of high-throughput automated methods used in early drug discovery research is also provided in this report. This review summarizes the challenges that are encountered during solubility measurement and the complexities that are often overlooked. Even though there is an advantage in using the amorphous form of a drug due to its higher solubility, measurement of its solubility with useful accuracy is still a practical problem. Therefore, this review provides recommendations of preferred methods and precautions in using these methods to determine the aqueous solubility of amorphous and crystalline new molecular entities, with emphasis on the physico-chemical characterization of the solid state of the test substance.

Key factors governing the reconstitution time of high concentration lyophilized protein formulations.

Lyophilized protein formulations containing highly concentrated proteins often have long and variable reconstitution times. Reconstitution time is dependent on a number of factors in a complex manner. Furthermore, factors influencing the reconstitution of partially crystalline cakes are reportedly different from those of amorphous cakes. The objectives of this work were to identify the key factors governing reconstitution and understand the mechanisms involved in reconstitution of both amorphous and partially crystalline cakes. Partial crystallinity in the final cake, larger pores and low "concentrated formulation viscosity" (i.e., viscosity near the surface of the dissolving cake) were identified as desirable characteristics for expediting reconstitution. Crystallinity and larger pores dramatically improved wettability and liquid penetration into partially crystalline cakes, ultimately resulting in well dispersed small pieces of partially dissolved cake. The smaller disintegrated cake pieces dissolved faster because of the increased surface area. The amorphous cakes exhibited poorer wettability than partially crystalline cakes. Moreover, the ability of the reconstitution fluid to penetrate the pores, and the resulting cake disintegration was much lower than that observed for partially crystalline cakes. In fact, for some of the amorphous cakes, the reconstitution fluid did not penetrate the cake at all. As a result, the undissolved intact cake or a large cake chunk floated on the reconstitution fluid amidst foam or bubbles generated during reconstitution. Dissolution of the floating cake appeared to proceed via gradual surface erosion where reconstitution time was found to be highly correlated with the viscosity near the surface of the dissolving cake solids. A higher viscosity prolonged reconstitution. Thus, both formulation and processing conditions can be tailored to achieve faster reconstitution. Including a crystallizable excipient proved to be beneficial. Incorporating an annealing step to facilitate crystallization of the crystallizable excipient and to promote larger pores was also found to be advantageous. A viscosity lowering excipient in the formulation could potentially be helpful but needs to be explored further.

Impact of formulation on the quality and stability of freeze-dried nanoparticles.

Freeze-drying is an effective approach to improve the long-term stability of nanomedicines. Lyoprotectants are generally considered as requisite excipients to ensure that the quality of nanoparticles is maintained throughout the freeze-drying process. However, depending on the type of nanoparticles, the needs for lyoprotectants or the challenges they face during freeze-drying may be different. In this study, we compared and identified the impact of freeze-drying on key characteristics of three types of nanoparticles: solid lipid nanoparticles (SLNs), polymeric nanoparticles (PNs), and liposomes. Sucrose, trehalose, and mannitol were added to nanoparticle suspensions before freeze-drying. The same conservative freeze-drying conditions with controlled ice nucleation at -8 °C were employed for all formulations. The collapse temperatures of nanoparticle formulations were found to be the same as those of the lyoprotectant added, except PN formulation. Likely the poly(vinyl alcohol) (PVA) in the formulation induced a higher collapse temperature and retardation of drying of PNs. Freeze-drying of both SLNs and liposomes without lyoprotectants increased particle size and polydispersity, which was resolved by adding amorphous disaccharides. Regardless of the addition of lyoprotectants, freeze-drying did not alter the size of PNs possibly due to the protection from PVA. However, lyoprotectants were still necessary to shorten the reconstitution time and reduce the residual moisture. In conclusion, different types of nanoparticles face distinct challenges for freeze-drying, and lyoprotectants differentially affect various stability and quality attributes of freeze-dried nanoparticles.

Solubility advantage of amorphous pharmaceuticals: II. Application of quantitative thermodynamic relationships for prediction of solubility enhancement in structurally diverse insoluble pharmaceuticals.

PURPOSE: To quantitatively assess the solubility advantage of amorphous forms of nine insoluble drugs with a wide range of physico-chemical properties utilizing a previously reported thermodynamic approach. METHODS: Thermal properties of amorphous and crystalline forms of drugs were measured using modulated differential calorimetry. Equilibrium moisture sorption uptake by amorphous drugs was measured by a gravimetric moisture sorption analyzer, and ionization constants were determined from the pH-solubility profiles. Solubilities of crystalline and amorphous forms of drugs were measured in de-ionized water at 25°C. Polarized microscopy was used to provide qualitative information about the crystallization of amorphous drug in solution during solubility measurement. RESULT: For three out the nine compounds, the estimated solubility based on thermodynamic considerations was within two-fold of the experimental measurement. For one compound, estimated solubility enhancement was lower than experimental value, likely due to extensive ionization in solution and hence its sensitivity to error in pKa measurement. For the remaining five compounds, estimated solubility was about 4- to 53-fold higher than experimental results. In all cases where the theoretical solubility estimates were significantly higher, it was observed that the amorphous drug crystallized rapidly during the experimental determination of solubility, thus preventing an accurate experimental assessment of solubility advantage. CONCLUSION: It has been demonstrated that the theoretical approach does provide an accurate estimate of the maximum solubility enhancement by an amorphous drug relative to its crystalline form for structurally diverse insoluble drugs when recrystallization during dissolution is minimal.

Reconstitution of Highly Concentrated Lyophilized Proteins: Part 1 Amorphous Formulations.

Long reconstitution times before patient administration remain an undesirable quality attribute for high concentration lyophilized protein formulations. In this study, 3 approaches were developed to study reconstitution behavior of lyophilized, amorphous cakes of a highly concentrated monoclonal antibody (mAb) by exploring their wetting, disintegration, and hydration behavior. As the mAb concentration increased from 0 to 83 mg/mL, reconstitution times were longer with poorer wetting, slower hydration, and disintegration rates. Furthermore, the effect of controlling ice nucleation temperature at -5 and -10°C during freezing followed by either conservative or aggressive drying conditions on the reconstitution times was explored in formulations containing 40 and 83 mg/mL mAb. Although no effect of either of the 2 processing conditions was noted at 40 mg/mL, aggressive drying led to faster reconstitution at both the nucleation temperatures with 83 mg/mL mAb. The present study combined with literature data suggests that below a protein-to-sugar ratio of 1, reconstitution was complete within 1 min, and when the ratio was greater than 1, the reconstitution times increased nonlinearly. Disintegration and hydration were determined to be the key mechanisms contributing to the complete reconstitution of the lyophilized, amorphous cakes of the highly concentrated mAb in vials.

Reconstitution Time for Highly Concentrated Lyophilized Proteins: Role of Formulation and Protein.

Lyophilized protein formulations containing highly concentrated proteins often have long reconstitution times. The goal was to understand the role of formulation in mediating the reconstitution time. Formulation variables such as % total solids, protein concentration, protein-to-sugar ratio, different proteins and inclusion of a crystallizable excipient were investigated for their effect on cake properties influencing reconstitution namely, cake wettability, penetration of reconstitution fluid into the cake, cake disintegration and cake porous structure. Additionally, several measures of viscosity were also evaluated for their effect on reconstitution time. Reconstitution time was primarily influenced by the "concentrated formulation viscosity" with negligible contributions from % total solids and protein concentration. "Concentrated formulation viscosity" was sensitive to both protein-to-sugar ratio and the protein itself. Partial crystallinity in the final cake also expedited reconstitution. Wettability, liquid penetration into the cake, cake disintegration tendency and cake porous structure were found to be invariant for amorphous cakes and did not correlate with reconstitution time. However, these properties were sensitive to the presence of crystallinity and resulted in faster reconstitution at least of the partially crystalline cakes. "Concentrated formulation viscosity" strongly correlated with reconstitution times of amorphous cakes, providing insights on the steps involved in the reconstitution of amorphous formulations.

Stability of Freeze-Dried Protein Formulations: Contributions of Ice Nucleation Temperature and Residence Time in the Freeze-Concentrate.

Controlling ice nucleation, at a fixed higher temperature, results in larger ice crystals, which can reduce the ice/freeze-concentrate interface area where proteins can adsorb and partially unfold. Moreover, limited work has been done to address any effects on short-term stability due to a slow ramp or long isothermal hold after the ice nucleation step. The objective was to evaluate the effect of the ice nucleation temperature and residence time in the freeze-concentrate on in-process or storage stability of representative proteins, human IgG, and recombinant human serum albumin. The results suggest a higher ice nucleation temperature can minimize aggregation of protein pharmaceuticals, which are labile at ice/aqueous interface. Apart from the ice nucleation step, the present study identified the residence time in the freeze-concentrate as the critical factor that influences protein stability post ice nucleation. At a temperature where enough mobility exists (i.e., above Tg' of the formulation), the long residence time in the freeze-concentrate can result in significant protein aggregation during the process. In addition to stability, the findings revealed that not only the ice nucleation temperature but also the thermal history of the formulation post ice nucleation defines the surface area of ice and the porous structure of the freeze-dried cake.

Nuances in the Calculation of Amorphous Solubility Enhancement Ratio.

The theoretical amorphous solubility enhancement ratio (Rs) can be calculated based on the free energy difference between amorphous and crystalline forms (ΔGx→a), using several experimentally determined input parameters. This work compares the various approaches for the calculation of Rs and explores the nuances associated with its calculation. The uncertainty of Rs values owing to experimental conditions (differential scanning calorimetry heating rates) used to measure the input parameters was determined for 3 drugs (indomethacin, itraconazole, and spironolactone). The calculated value of Rs was most influenced by the measurement of heat of fusion. The range in values of Rs using the various equations in the literature was within the calculated uncertainty of the theoretical Rs value. Still, all equations appear to overpredict the experimental value of Rs, sometimes by more than a factor of 5, when an experimental value is attainable. Methods for the calculation of ΔGx→a for molecules undergoing additional phase transitions (other than glass transition and melting) were developed, employing itraconazole as a model drug. In addition, the influences of enthalpy relaxation and entropy of mixing for racemic compounds on Rs were also considered. These additional corrections improved agreement between theoretical and experimental Rs.

Study of the individual contributions of ice formation and freeze-concentration on isothermal stability of lactate dehydrogenase during freezing.

The objective of this study was to determine the individual contributions of ice formation, solute concentration, temperature, and time, to irreversible protein denaturation during freezing. A temperature-step approach was used to study isothermal degradation of frozen lactate dehydrogenase (LDH). The freeze-concentrate composition was determined using differential scanning calorimetry to enable preparation of solutions, without ice, of the same concentration as the freeze-concentrate, and thereby determine the role of the freeze-concentrate composition on LDH degradation. Both stabilizers employed in the study, hydroxyethyl starch and sucrose, conferred cryoprotection on LDH. While LDH stability was lower at 1.50-3.25% w/v sucrose than in the absence of sucrose, cryoprotection was restored at higher sucrose concentrations. pH shift during freezing, degree of supercooling, and excipient impurities were ruled out as causes for unusual LDH stability behavior at lower sucrose concentrations. Specific surface area measurements of the freeze-dried cakes showed that the ice surface area increased with an increase in sucrose concentration. No LDH degradation occurred in concentrated solutions, without ice, at the same composition as the freeze-concentrate in frozen systems where massive degradation was documented. Thus, ice formation is the critical destabilizing factor during freezing of LDH in sucrose:citrate buffer systems.

Amorphization alone does not account for the enhancement of solubility of drug co-ground with silicate: the case of indomethacin.

The solubility advantage of indomethacin amorphized by co-grinding with Neusilin US2 in various media was investigated. Physical mixtures of gamma-indomethacin and Neusilin US2 (in the ratios 1:1, 1:4 and 1:5) were amorphized at room temperature employing 75% RH in a porcelain jar mill using zirconia balls. The crystallinity of the samples was determined using ATR-FTIR and PXRD. The solubility and dissolution profiles of co-ground powders and crystalline counterparts were evaluated in 0.1 N HCl, water and phosphate buffer (pH 6.8) in a USP type II dissolution apparatus at 250 rpm and 37 degrees C. Very high concentrations of dissolved indomethacin as compared to the solubility of gamma-indomethacin (approximately 500 times in water and approximately 3.7 times in phosphate buffer) were attained. However, the presence of other polymorphs detected by PXRD and a change in the pH of the medium made interpretation of the results difficult. In 0.1 N HCl the solubility (i.e., the peak in a concentration versus time plot) of the amorphized drug in a 1:5 ratio with Neusilin increased to 109 times the solubility of crystalline gamma-indomethacin alone. An increase in amount of drug and Neusilin in the same ratio added to the dissolution medium also increased peak and plateau dissolution concentrations. The presence of silicic acid and ions (Mg(2+) and Al(3+)) in the dissolution media were found to cause the increase in the plateau concentration of indomethacin. Amorphization alone does not account for all of the dissolution enhancement; acidity, ions, and silicic acid are major contributors to dissolution enhancement.