Hydrogen Bonding Interactions in Amorphous Indomethacin and Its Amorphous Solid Dispersions with Poly(vinylpyrrolidone) and Poly(vinylpyrrolidone-co-vinyl acetate) Studied Using (13)C Solid-State NMR.

Hydrogen bonding interactions in amorphous indomethacin and amorphous solid dispersions of indomethacin with poly(vinylpyrrolidone), or PVP, and poly(vinylpyrrolidone-co-vinyl acetate), or PVP/VA, were investigated quantitatively using solid-state NMR spectroscopy. Indomethacin that was (13)C isotopically labeled at the carboxylic acid carbon was used to selectively analyze the carbonyl region of the spectrum. Deconvolution of the carboxylic acid carbon peak revealed that 59% of amorphous indomethacin molecules were hydrogen bonded through carboxylic acid cyclic dimers, 15% were in disordered carboxylic acid chains, 19% were hydrogen bonded through carboxylic acid and amide interactions, and the remaining 7% were free of hydrogen bonds. The standard dimerization enthalpy and entropy of amorphous indomethacin were estimated to be -38 kJ/mol and -91 J/(mol · K), respectively, using polystyrene as the "solvent". Polymers such as PVP and PVP/VA disrupted indomethacin self-interactions and formed hydrogen bonds with the drug. The carboxylic acid dimers were almost completely disrupted with 50% (wt) of PVP or PVP/VA. The fraction of disordered carboxylic acid chains also decreased as the polymer content increased. The solid-state NMR results were compared with molecular dynamics (MD) simulations from the literature. The present work highlights the potential of (13)C solid-state NMR to detect and quantify various hydrogen bonded species in amorphous solid dispersions as well as to serve as an experimental validation of MD simulations.

Release, partitioning, and conjugation stability of doxorubicin in polymer micelles determined by mechanistic modeling.

PURPOSE: To better understand the mechanistic parameters that govern drug release from polymer micelles with acid-labile linkers. METHODS: A mathematical model was developed to describe drug release from block copolymer micelles composed of a poly(ethylene glycol) shell and a poly(aspartate) core, modified with drug binding linkers for pH-controlled release [hydrazide (HYD), aminobenzoate-hydrazide (ABZ), or glycine-hydrazide (GLY)]. Doxorubicin (Dox) was conjugated to the block copolymers through acid-labile hydrazone bonds. The polymer drug conjugates were used to prepare three polymer micelles (HYD-M, ABZ-M, and GLY-M). Drug release studies were performed to identify the factors governing pH-sensitive release of Dox. The effect of prolonged storage of copolymer material on release kinetics was also observed. RESULTS: Biphasic drug release kinetics were observed for all three micelle formulations. The developed model was able to quantify observed release kinetics upon the inclusion of terms for unconjugated Dox and two populations of conjugated Dox. Micelle/water partitioning of Dox was also incorporated into the model and found significant in all micelles under neutral conditions but reduced under acidic conditions. The drug binding linker played a major role in drug release as the extent of Dox release at specific time intervals was greater at pH 5.0 than at pH 7.4 (HYD-M > ABZ-M > GLY-M). Mathematical modeling was also able to correlate changes in release kinetics with the instability of the hydrazone conjugation of Dox during prolonged storage. CONCLUSION: These results illustrate the potential utility of mechanistic modeling to better assess release characteristics intrinsic to a particular drug/nanoparticle system.

Dynamic, nonsink method for the simultaneous determination of drug permeability and binding coefficients in liposomes.

Drug release from liposomal formulations is governed by a complex interplay of kinetic (i.e., drug permeability) and thermodynamic factors (i.e., drug partitioning to the bilayer surface). Release studies under sink conditions that attempt to mimic physiological conditions are insufficient to decipher these separate contributions. The present study explores release studies performed under nonsink conditions coupled with appropriate mathematical models to describe both the release kinetics and the conditions in which equilibrium is established. Liposomal release profiles for a model anticancer agent, topotecan, under nonsink conditions provided values for both the first-order rate constant for drug release and the bilayer/water partition coefficient. These findings were validated by conducting release studies under sink conditions via dynamic dialysis at the same temperature and buffer pH. A nearly identical rate constant for drug release could be obtained from dynamic dialysis data when appropriate volume corrections were applied and a mechanism-based mathematical model was employed to account for lipid bilayer binding and dialysis membrane transport. The usefulness of the nonsink method combined with mathematical modeling was further explored by demonstrating the effects of topotecan dimerization and bilayer surface charge potential on the bilayer/water partition coefficient at varying suspension concentrations of lipid and drug.

Effect of precipitation inhibitors on indomethacin supersaturation maintenance: mechanisms and modeling.

This study quantitatively explores the mechanisms underpinning the effects of model pharmaceutical polymeric precipitation inhibitors (PPIs) on the crystal growth and, in turn, maintenance of supersaturation of indomethacin, a model poorly water-soluble drug. A recently developed second-derivative UV spectroscopy method and a first-order empirical crystal growth model were used to determine indomethacin crystal growth rates in the presence of model PPIs. All three model PPIs including HP-ß-CD, PVP, and HPMC inhibited indomethacin crystal growth at both high and low degrees of supersaturation (S). The bulk viscosity changes in the presence of model PPIs could not explain their crystal growth inhibitory effects. At 0.05% w/w, PVP (133-fold) and HPMC (28-fold) were better crystal growth inhibitors than HP-ß-CD at high S. The inhibitory effect of HP-ß-CD on the bulk diffusion-controlled indomethacin crystal growth at high S was successfully modeled using reactive diffusion layer theory, which assumes reversible complexation in the diffusion layer. Although HP-ß-CD only modestly inhibited indomethacin crystal growth at either high S (∼15%) or low S (∼2-fold), the crystal growth inhibitory effects of PVP and HPMC were more dramatic, particularly at high S (0.05% w/w). The superior crystal growth inhibitory effects of PVP and HPMC as compared with HP-ß-CD at high S were attributed to a change in the indomethacin crystal growth rate-limiting step from bulk diffusion to surface integration. Indomethacin crystal growth inhibitory effects of all three model PPIs at low S were attributed to retardation of the rate of surface integration of indomethacin, a phenomenon that may reflect the adsorption of PPIs onto the growing crystal surface. The quantitative approaches outlined in this study should be useful in future studies to develop tools to predict supersaturation maintenance effects of PPIs.

Molecular dynamics simulation of amorphous hydroxypropyl-methylcellulose acetate succinate (HPMCAS): polymer model development, water distribution, and plasticization.

Molecular models for HPMCAS polymer have been developed for molecular dynamics (MD) simulation that attempt to mimic the complex substitution patterns in HPMCAS observed experimentally. These molecular models were utilized to create amorphous HPMCAS solids by cooling of the polymeric melts at different water contents to explore the influence of water on molecular mobility, which plays a critical role in stability and drug release from HPMCAS-based solid matrices. The densities found for the simulated amorphous HPMCAS were 1.295, 1.287, and 1.276 g/cm(3) at 0.7, 5.7, and 13.2% w/w water, indicating swelling of the polymer with increasing water content. These densities compare favorably with the experimental density of 1.285 g/cm(3) for commercial HPMCAS-(AQOAT AS-MF) supporting the present HPMCAS models as a realistic representation of amorphous HPMCAS solids. Water molecules were observed to be mostly isolated from each other at a low water content (0.7% w/w), while clusters or strands of water were pervasive and broadly distributed in size at 13.2% w/w water. The average number of first-shell water molecules (n(w)) increased from 0.17 to 3.5, though the latter is still far below that (8.9) expected for the onset of a separate water phase. Increasing water content from 0.7 to 13.2% w/w was found to reduce the T(g) by ~81 K, similar to experimental observations. Plasticization with increasing water content resulted in increasing polymer mobility and water diffusivity. From 0.7 to 13.2% w/w water, the apparent water diffusivity increased from 1.1 × 10(-9) to 7.0 × 10(-8) cm(2)/s, though non-Einsteinian behavior persisted at all water contents explored. This and the water trajectories in the polymers suggest that water diffusion at 0.7% w/w water follows a "hopping" mechanism. At a higher water content (13.2% w/w) water diffusion follows dual diffusive processes: (1) fast water motions within water clusters; and (2) slower diffusion through the more rigid polymer matrix.

Molecular dynamics simulation of amorphous indomethacin.

Molecular dynamics (MD) simulations have been conducted using an assembly consisting of 105 indomethacin (IMC) molecules and 12 water molecules to investigate the underlying dynamic (e.g., rotational and translational diffusivities and conformation relaxation rates) and structural properties (e.g., conformation, hydrogen-bonding distributions, and interactions of water with IMC) of amorphous IMC. These properties may be important in predicting physical stability of this metastable material. The IMC model was constructed using X-ray diffraction data with the force-field parameters mostly assigned by analogy with similar groups in Amber-ff03 and atomic charges calculated with the B3LYP/ccpVTZ30, IEFPCM, and RESP models. The assemblies were initially equilibrated in their molten state and cooled through the glass transition temperature to form amorphous solids. Constant temperature dynamic runs were then carried out above and below the T(g) (i.e., at 600 K (10 ns), 400 K (350 ns), and 298 K (240 ns)). The density (1.312 ± 0.003 g/cm(3)) of the simulated amorphous solid at 298 K was close to the experimental value (1.32 g/cm(3)) while the estimated T(g) (384 K) was ~64 degrees higher than the experimental value (320 K) due to the faster cooling rate. Due to the hindered rotation of its amide bond, IMC can exist in different diastereomeric states. Different IMC conformations were sufficiently sampled in the IMC melt or vapor, but transitions occurred rarely in the glass. The hydrogen-bonding patterns in amorphous IMC are more complex in the amorphous state than in the crystalline polymorphs. Carboxylic dimers that are dominant in α- and γ-crystals were found to occur at a much lower probability in the simulated IMC glasses while hydrogen-bonded IMC chains were more easily identified patterns in the simulated amorphous solids. To determine molecular diffusivity, a novel analytical method is proposed to deal with the non-Einsteinian behavior, in which the temporal evolution of the apparent diffusivity D(t) is described by a relaxation model such as the KWW function and extrapolated to infinite time. The diffusion coefficient found for water diffusing in amorphous indomethacin at 298 K (2.7 × 10(-9) cm(2)/s) compares favorably to results obtained in experimental IMC glasses (0.9-2.0 × 10(-9) cm(2)/s) and is mechanistically associated with ß-relaxation processes that are dominant in sub-T(g) glasses.

Determination of drug release kinetics from nanoparticles: overcoming pitfalls of the dynamic dialysis method.

Dynamic dialysis is one of the most common methods for the determination of release kinetics from nanoparticle drug delivery systems. Drug appearance in the "sink" receiver compartment is a consequence of release from the nanoparticles into the dialysis chamber followed by diffusion across the dialysis membrane. This dual barrier nature inherent in the method complicates data interpretation and may lead to incorrect conclusions regarding nanoparticle release half-lives. Although the need to consider the barrier properties of the dialysis membrane has long been recognized, there is insufficient quantitative appreciation for the role of the driving force for drug transport across that membrane. Reversible nanocarrier binding of the released drug reduces the driving force for drug transport across the dialysis membrane leading to a slower overall apparent release rate. This may lead to the conclusion that a given nanoparticle system will provide a sustained release in vivo when it will not. This study demonstrates these phenomena using model lipophilic drug-loaded liposomes varying in lipid composition to provide variations in bilayer permeability and membrane binding affinities. Model simulations of liposomal transport as measured by dynamic dialysis were conducted to illustrate the interplay between the liposome concentration, membrane/water partition coefficient, and the apparent release rate. Reliable determination of intrinsic liposomal bilayer permeability coefficients for lipophilic drugs by dynamic dialysis requires validation of drug release kinetics at varying nanoparticle concentration and the determination of membrane binding coefficients along with appropriate mechanism-based mathematical modeling to ensure the reliability and proper interpretation of the data.

Bilayer composition, temperature, speciation effects and the role of bilayer chain ordering on partitioning of dexamethasone and its 21-phosphate.

PURPOSE: Models to predict membrane-water partition coefficients (Kp) as a function of drug structure, membrane composition, and solution properties would be useful. This study explores the partitioning of dexamethasone (Dex) and its ionizable 21-phosphate (Dex-P) in liposomes varying in acyl chain length, physical state, and pH. METHODS: DMPC:mPEG DMPE, DPPC:mPEG DPPE, and DSPC:mPEG DSPE (95:5 mol%) liposomes were prepared by thin film hydration. Kp values for Dex and Dex-P were determined from pH 1.5-8 by equilibrium dialysis and equilibrium solubility (Dex). RESULTS: Dex Kp values at 25°C were 705 ± 24, 106 ± 11, and 58 ± 9 in DMPC, DPPC, and DSPC, increasing to 478 ± 20 in DPPC liposomes at 45°C. Both neutral and anionic species contributed to the Kp of Dex-P versus solution pH (1.5-8). A linear correlation was found between the natural logarithm of Kp and the inverse of bilayer free surface area (1/afree) where afree is a parameter reflecting chain ordering that depends on bilayer composition and temperature. CONCLUSIONS: Models of the pH dependence of partitioning of ionizable compounds must include contributions of both neutral and ionized species. Bilayer free surface area may be an important variable to predict Kp of drug molecules versus lipid composition and temperature.

Kinetics and mechanisms of deamidation and covalent amide-linked adduct formation in amorphous lyophiles of a model asparagine-containing Peptide.

PURPOSE: Asparagine containing peptides and proteins undergo deamidation via a succinimide intermediate. This study examines the role of the succinimide in the formation of covalent, amide-linked adducts in amorphous peptide formulations. METHODS: Stability studies of a model peptide, Gly-Phe-L-Asn-Gly, were performed in lyophiles containing an excess of Gly-Val at 'pH' 9.5 and 40°C/40% RH. Reactant disappearance and the formation of ten different degradants were monitored by HPLC. Mechanism-based kinetic models were used to generate rate constants from the concentration vs. time profiles. RESULTS: Deamidation of Gly-Phe-L-Asn-Gly in lyophiles resulted in L- and D-aspartyl and isoaspartyl-containing peptides and four amide-linked adducts between the succinimide and Gly-Val. The kinetic analysis demonstrated competition between water and terminal amino groups in Gly-Val for the succinimide. The extent of covalent adduct formation was dependent on dilution effects due to its second order rate law. CONCLUSION: The cyclic imide formed during deamidation of asparagine containing peptides in lyophiles can also lead to covalent adducts due to reaction with other neighboring peptides. A reaction model assuming a central role for the succinimide in the formation both hydrolysis products and covalent adducts was quantitatively consistent with the kinetic data. This mechanism may contribute to the presence of covalent, non-reducible aggregates in lyophilized peptide formulations.

Pharmacokinetic modeling to assess factors affecting the oral bioavailability of the lactone and carboxylate forms of the lipophilic camptothecin analogue AR-67 in rats.

PURPOSE: Camptothecin analogues are anticancer drugs effective when dosed in protracted schedules. Such treatment is best suited for oral formulations. AR-67 is a novel lipophilic analogue with potent efficacy in preclinical models. Here we assessed factors that may influence its oral bioavailability in rats. METHODS: Plasma pharmacokinetic (PK) studies were conducted following administration of AR-67 lactone or carboxylate doses alone or after pre-dosing with inhibitors of the efflux transporters P-gp and Bcrp. A population PK model that simultaneously fitted to oral and intravenous data was used to estimate the bioavailability (F) and clearance of AR-67. RESULTS: An inverse Gaussian function was used as the oral input into the model and provided the best fits. Covariate analysis showed that the bioavailability of the lactone, but not its clearance, was dose dependent. Consistent with this observation, the bioavailability of AR-67 increased when animals were pretreated orally with GF120918 or Zosuquidar. CONCLUSION: Absorption of AR-67 is likely affected by solubility of its lactone form and interaction with efflux pumps in the gut. AR-67 appears to be absorbed as the lactone form, most likely due to gastric pH favoring its formation and predominance. F increased at higher doses suggesting saturation of efflux mechanisms.

Genome sequence of the plant-pathogenic bacterium Dickeya dadantii 3937.

Dickeya dadantii is a plant-pathogenic enterobacterium responsible for the soft rot disease of many plants of economic importance. We present here the sequence of strain 3937, a strain widely used as a model system for research on the molecular biology and pathogenicity of this group of bacteria.

An atomic and molecular view of the depth dependence of the free energies of solute transfer from water into lipid bilayers.

Molecular interactions and orientations responsible for differences in 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) bilayer partitioning of three structurally related drug-like molecules (4-ethylphenol, phenethylamine, and tyramine) were investigated. This work is based on previously reported molecular dynamics (MD) simulations that determined their transverse free energy profiles across the bilayer. Previously, the location where the transfer free energy of the three solutes is highest, which defines the barrier domain for permeability, was found to be the bilayer center, while the interfacial region was found to be the preferred binding region. Contributions of the amino (NH2) and hydroxyl (OH) functional groups to the transfer free energies from water to the interfacial region were found to be very small both experimentally and by MD simulation, suggesting that the interfacial binding of these solutes is hydrophobically driven and occurs with minimal loss of hydrogen-bonding interactions of the polar functional groups which can occur with either water or phospholipid head groups. Therefore, interfacial binding is relatively insensitive to the number or type of polar functional groups on the solute. In contrast, the relative solute free energy in the barrier domain is highly sensitive to the number of polar functional groups on the molecule. The number and types of hydrogen bonds formed between the three solutes and polar phospholipid atoms or with water molecules were determined as a function of solute position in the bilayer. Minima were observed in the number of hydrogen bonds formed by each solute at the center of the bilayer, coinciding with a decrease in the number of water molecules in DOPC as a function of distance away from the interfacial region. In all regions, hydrogen bonds with water molecules account for the majority of hydrogen-bonding interactions observed for each solute. Significant orientational preferences for the solutes are evident in certain regions of the bilayer (e.g., within the ordered chain region and near the interfacial region 20-25 Å from the bilayer center). The preferred orientations are those that preserve favorable molecular interactions for each solute, which vary with the solute structure.

Factors affecting the in vivo lactone stability and systemic clearance of the lipophilic camptothecin analogue AR-67.

PURPOSE: The narrow efficacy-toxicity window of anticancer agents necessitates understanding of factors contributing to their disposition. This is especially true for camptothecins as they exist in the lactone and carboxylate forms with each moiety differentially interacting with efflux or uptake transporters. Here we determined the disposition of the lactone and carboxylate forms of AR-67, a 3(rd) generation camptothecin analogue. METHODS: Pharmacokinetic studies were conducted in rats given intravenous boluses of AR-67 lactone or carboxylate with or without pharmacologic inhibitor pretreatment (GF120918 or rifampin). Pharmacokinetic modeling was used to estimate clearances, while simulations assessed the impact of clearance changes on overall AR-67 exposure. RESULTS: Our modeling showed that carboxylate clearance was 3.5-fold higher than that of the lactone. GF120918 decreased lactone clearance only, but rifampin decreased both lactone and carboxylate clearances. Simulations showed that decreasing carboxylate clearance, which controls the overall drug disposition, leads to significant increase in AR-67 exposure. CONCLUSION: The apparent in vivo blood stability of AR-67 is partly dependent on the increased carboxylate clearance. This may have clinical implications for populations with single nucleotide polymorphisms that impair the function of uptake transporter genes (e.g., SLCO1B1), which are potentially responsible for AR-67 carboxylate clearance.

Enteropathogen Resource Integration Center (ERIC): bioinformatics support for research on biodefense-relevant enterobacteria.

ERIC, the Enteropathogen Resource Integration Center (www.ericbrc.org), is a new web portal serving as a rich source of information about enterobacteria on the NIAID established list of Select Agents related to biodefense-diarrheagenic Escherichia coli, Shigella spp., Salmonella spp., Yersinia enterocolitica and Yersinia pestis. More than 30 genomes have been completely sequenced, many more exist in draft form and additional projects are underway. These organisms are increasingly the focus of studies using high-throughput experimental technologies and computational approaches. This wealth of data provides unprecedented opportunities for understanding the workings of basic biological systems and discovery of novel targets for development of vaccines, diagnostics and therapeutics. ERIC brings information together from disparate sources and supports data comparison across different organisms, analysis of varying data types and visualization of analyses in human and computer-readable formats.

Effects of Molecular Interactions on Miscibility and Mobility of Ibuprofen in Amorphous Solid Dispersions With Various Polymers.

Hydrogen bonds (HBs) in amorphous solid dispersions may influence physical stability through effects on both drug miscibility and mobility. Amorphous solid dispersions containing the HB-donor ibuprofen (IBP) alone or with one of four model polymers (poly(vinyl pyrrolidone) [PVP], poly(vinyl pyrrolidone/vinyl acetate) [PVP/VA], poly(vinyl acetate) [PVA], or polystyrene [PST]) were monitored by molecular dynamics simulation. HB distributions and contributions of electrostatic, van der Waals, and internal interactions to miscibility and mobility were analyzed versus drug concentration. The probability of IBP-IBP HBs decreases markedly (0.6→0.0) with dilution (100→10% drug) in PVP due to IBP-PVP HBs while dilution in the nonpolar PST has a more modest effect on IBP-IBP HB probability (0.6→0.3). Concentration-dependent Flory-Huggins interaction parameters (χ) were determined to assess drug-polymer miscibility. χIBP-PVP values were -0.9 to -1.8 with a plateau near 50% w/w PVP, whereas χIBP-PST fluctuated near zero (-0.1 to 0.3), suggesting that IBP is more soluble in PVP than in PST. χIBP-polymer values in polymers varying in pyrrolidone/acetate composition were in the order PVP (most favorable) > PVP/VA > PVA (least favorable). Decreased local mobility of IBP measured by the atomic fluctuation correlates with more IBP-PVP HBs with increasing PVP content. The opposite trend in IBP-PST may arise from IBP-IBP HB disruption on dilution.

Fifty-Eight Years and Counting: High-Impact Publishing in Computational Pharmaceutical Sciences and Mechanism-Based Modeling.

With this issue of the Journal of Pharmaceutical Sciences, we celebrate the nearly 6 decades of contributions to mechanistic-based modeling and computational pharmaceutical sciences. Along with its predecessor, The Journal of the American Pharmaceutical Association: Scientific Edition first published in 1911, JPharmSci has been a leader in the advancement of pharmaceutical sciences beginning with its inaugural edition in 1961. As one of the first scientific journals focusing on pharmaceutical sciences, JPharmSci has established a reputation for publishing high-quality research articles using computational methods and mechanism-based modeling. The journal's publication record is remarkable. With over 15,000 articles, 3000 notes, and more than 650 reviews from industry, academia, and regulatory agencies around the world, JPharmSci has truly been the leader in advancing pharmaceutical sciences.

Frenolicin B Targets Peroxiredoxin 1 and Glutaredoxin 3 to Trigger ROS/4E-BP1-Mediated Antitumor Effects.

Peroxiredoxin 1 (Prx1) and glutaredoxin 3 (Grx3) are two major antioxidant proteins that play a critical role in maintaining redox homeostasis for tumor progression. Here, we identify the prototypical pyranonaphthoquinone natural product frenolicin B (FB) as a selective inhibitor of Prx1 and Grx3 through covalent modification of active-site cysteines. FB-targeted inhibition of Prx1 and Grx3 results in a decrease in cellular glutathione levels, an increase of reactive oxygen species (ROS), and concomitant inhibition of cancer cell growth, largely by activating the peroxisome-bound tuberous sclerosis complex to inhibit mTORC1/4E-BP1 signaling axis. FB structure-activity relationship studies reveal a positive correlation between inhibition of 4E-BP1 phosphorylation, ROS-mediated cancer cell cytotoxicity, and suppression of tumor growth in vivo. These findings establish FB as the most potent Prx1/Grx3 inhibitor reported to date and also notably highlight 4E-BP1 phosphorylation status as a potential predictive marker in response to ROS-based therapies in cancer.

What determines drug solubility in lipid vehicles: is it predictable?

Lipid-based drug delivery systems are of increasing interest to the pharmaceutical scientist because of their potential to solubilize drug molecules that may be otherwise difficult to develop. The ability to predict lipid solubility is an important step in being able to identify the right excipients to solubilize and formulate drugs in lipid formulations. However, predicting lipid solubility is complicated by the fact that interfacial effects may play a dominant role in these mixtures and the solubility may be affected by the microstructure (microemulsions, emulsions, oily solutions, etc.), as well as by the physicochemical properties of the oil, surfactant, co-solvent, and the drug. This review illustrates the fundamental factors that govern solubility in lipid mixtures and discusses models built at varying levels of sophistication to estimate the solubility. Examples from the literature are presented that demonstrate the application of these models, how their choice is related to the drug/lipid employed, and the challenges involved in solubility prediction. New data on the role water plays in altering lipid solubility, not only through its interaction with the solute, but also by changing the structure of lipids by promoting lipid organization are highlighted. The available data demonstrate that a rational understanding of solubilization in lipids is a worthwhile pursuit and models to predict at least the relative solubility from chemical structure have potential. Prediction of absolute solubility is more difficult as it requires knowledge of the drug's escaping tendency from the crystalline state. In recent years, it has become amply clear that for polar solutes, specific interactions are a critical factor governing solubility. Methods that can better take into account the specific as well as non-specific interactions between the solute and solvent, and the lipid microstructure, hold considerable promise for the future.

Liposomal delivery of hydrophobic weak acids: enhancement of drug retention using a high intraliposomal pH.

Clinical development of highly potent lipophilic neutral camptothecins has been impeded by the poor solubility, stability, and nonspecific toxicity of these compounds. Liposomal encapsulation offers a promising formulation route for tumor site-specific delivery of these novel drug candidates. However, the development of formulation strategies for liposomal loading and retention of hydrophobic drugs such as the neutral camptothecins has been lacking. In the studies presented here, we explored the potential of a trans-bilayer pH gradient strategy for prolonging the liposome retention of DB-67, a novel lipophilic camptothecin that can undergo lactone ring-opening to form a hydrophobic weak acid. The liposome membrane permeability of DB-67 was obtained as a function of pH in aqueous buffers. A permeability model was developed and liposome membrane permeability was shown to be controlled by the fraction of unbound neutral lactone entrapped in the vesicles. Liposome membrane permeability of DB-67 was also studied under physiological conditions. The high membrane partitioning of DB-67 in the intraliposomal microenvironment was found to shift the equilibrium between lactone and carboxylate towards the lactone species resulting in a faster than desired drug release under physiological conditions. The effectiveness of the pH gradient strategy was further reduced under physiological conditions by the rapid loss of trans-membrane pH gradients due to CO(2) uptake. Simulations were conducted to explore the role of membrane binding, intravesicular pH, and carbonate buffer concentration in successful utilization of the pH gradient strategy for hydrophobic weak acids.

Influence of intravesicular pH drift and membrane binding on the liposomal release of a model amine-containing permeant.

Accurate determination of intrinsic permeability coefficients is critical to the development of structure-permeability relationships and liposomal delivery systems. The apparent release rate of a drug from liposomes may reflect not only its intrinsic permeability coefficient and barrier properties but also a variety of underlying equilibria including drug ionization, membrane binding or complexation, and kinetic processes such as buffer exchange. Additionally, transport of ionizable drugs that are initially at high concentrations in liposomes can generate or dissipate pH gradients across the barrier causing deviations from classical pH-permeability profiles. In this study, the liposomal release of a model amine (tyramine) is determined as a function of drug loading, intravesicular pH, and buffer composition. Kinetic models are derived to study effects of such equilibria (e.g., ionization, membrane binding) and kinetic processes (e.g., pH drift and acid/base carriers). All equilibrium constants needed for the models were independently measured and used. The barrier properties of the lipid bilayers under the experimental conditions were assessed by monitoring the transport of mannitol and bretylium as a function of pH. A corrected intrinsic permeability coefficient of 0.04 cm/s was in close agreement with the value predicted from the barrier domain model for bilayer permeability, suggesting that all perturbing factors were properly addressed.