Aggregate Determination by Permeation Technique.

Permeation technique is used to study molecular aggregation in aqueous solutions including formation of cyclodextrin guest/host aggregates. Since only guest molecules, host molecules and guest/host aggregates that are smaller than the pore size of a given semipermeable membrane are able to permeate through the membrane, negative deviation of permeation profiles indicates formation of guest/host aggregates or self-aggregates. This chapter describes how the method is used to detect formation of nano-sized aggregates and to determine the critical aggregation concentration (cac) from permeation profiles of a guest molecule.

Characterization and Evaluation of Ternary Complexes of Ascorbic Acid with γ-Cyclodextrin and Poly(vinyl Alcohol).

Ascorbic acid (AA) is a general antioxidant used in aqueous pharmaceutical formulations. However, in aqueous solutions, AA is unstable and easily oxidized when exposed to air, light and/or heat. Cyclodextrins are well known for their ability to form inclusion complexes with various compounds to improve their solubility and stability. Previous studies demonstrate that cyclodextrins preserve the antioxidant capacity of AA but data for γ-cyclodextrin (γCD) have not been reported. Poly(vinyl alcohol) (PVA) is a hydrophilic polymer widely used as a drug matrix in various pharmaceutical fields, but its application for drug stabilization is limited. This study aimed to investigate the protective ability of γCD on AA through the formation of ternary complexes with PVA. Binary (i.e., AA/γCD, AA/PVA and γCD/PVA) and ternary (i.e., AA/γCD/PVA) complexes were first confirmed. It was reported that those complexes were formed through interactions between the heterocyclic ring of AA, hydroxyl group of PVA and hydrophobic cavity of γCD. The hydrodynamic diameter of complexes was then studied. It was found that the diameter of γCD/PVA complexes increased with respect to the concentration of γCD. Higher γCD concentrations also resulted in increasing hydrodynamic diameters of the ternary complex. The presence of AA in ternary complexes interfered with the aggregation tendency of γCD/PVA binary complexes. Furthermore, the antioxidant capacity of AA in binary and ternary complexes was investigated. It was found that the presence of γCD preserved the antioxidant activity of AA, whereas PVA showed a contrasting effect. The influence of γCD and PVA concentration on antioxidant capacity was then studied through central composite design (CCD). Even though the concentration of γCD significantly affected the inhibition efficiency of the ternary complex, the insignificant influence of PVA could not be ignored. A promising protective ternary complex should consist of an optimized concentration of PVA and a high concentration of γCD.

Self-association of cyclodextrins and cyclodextrin complexes in aqueous solutions.

Cyclodextrins (CDs) are oligosaccharides that self-assemble in aqueous solutions to form transient clusters, nanoparticles and small microparticles. The critical aggregation concentration (cac) of the natural αCD, ßCD and γCD in pure aqueous solutions was estimated to be 25, 8 and 9 mg/ml, respectively. The cac of 2-hydroxypropyl-ß-cyclodextrin (HPßCD), that consists of mixture of isomers, was estimated to be significantly higher or 118 mg/ml. Addition of chaotropic agents (i.e. that disrupts non-covalent bonds such as hydrogen bonds) to the aqueous media increases the cac. Formation of drug/CD complexes can increase or decrease the cac. Due to the transient nature of the CD clusters and nanoparticles they can be difficult to detect and their presence is frequently ignored. However, they have profound effect on the physiochemical properties of CDs and their pharmaceutical applications. For example, the values of stability constants of drug/CD complexes can be both concentration dependent and method dependent. Like in the case of micelles water-soluble polymers can enhance the solubilizing effect of CDs. Also, formation of drug/CD complex nanoparticles appears to increase the ability of CDs to enhance drug delivery through some mucosal membranes.

Solubility of Cyclodextrins and Drug/Cyclodextrin Complexes.

Cyclodextrins (CDs), a group of oligosaccharides formed by glucose units bound together in a ring, show a promising ability to form complexes with drug molecules and improve their physicochemical properties without molecular modifications. The stoichiometry of drug/CD complexes is most frequently 1:1. However, natural CDs have a tendency to self-assemble and form aggregates in aqueous media. CD aggregation can limit their solubility. Through derivative formation, it is possible to enhance their solubility and complexation capacity, but this depends on the type of substituent and degree of substitution. Formation of water-soluble drug/CD complexes can increase drug permeation through biological membranes. To maximize drug permeation the amount of added CD into pharmaceutical preparation has to be optimized. However, solubility of CDs, especially that of natural CDs, is affected by the complex formation. The presence of pharmaceutical excipients, such as water-soluble polymers, preservatives, and surfactants, can influence the solubilizing abilities of CDs, but this depends on the excipients' physicochemical properties. The competitive CD complexation of drugs and excipients has to be considered during formulation studies.

γ-Cyclodextrin.

γ-Cyclodextrin (γCD) is a cyclic oligosaccharide formed by bacterial digestion of starch and used as solubilizing agent and stabilizer in a variety of pharmaceutical and food products. γCD is a large (molecular weight 1297Da) hydrophilic molecule that does not readily permeate biological membranes and is rapidly digested by bacteria in the gastrointestinal tract. In humans γCD is metabolized by α-amylase that is found in, for example, saliva, bile fluid and tears. Thus, bioavailability of γCD is negligible. Also, γCD is readily excreted unchanged in the urine after parenteral administration. Like other cyclodextrins, γCD can form water-soluble inclusion complexes with many poorly-soluble compounds. In comparison with the natural αCD and ßCD, γCD has the largest hydrophobic cavity, highest water solubility and the most favorable toxicological profile. The focus of this review is production, physiochemical properties, pharmacokinetics, toxicity and applications of γCD and its derivatives. Also, the aggregation behavior of γCD in aqueous media is discussed.

The self-assemble of natural cyclodextrins in aqueous solutions: Application of miniature permeation studies for critical aggregation concentration (cac) determinations.

Permeation techniques can be applied to determine the critical aggregation concentration (cac) of natural cyclodextrins (CDs) in aqueous solutions although the method is both laborious and time consuming. In the present study, the permeation technique was modified and the influence of osmotic pressure, sampling time, CD concentration and molecular weight-cut off (MWCO) of the membrane were investigated in two different permeation units, that is Franz diffusion cells and Slide-A-Lyzer™ MINI Dialysis. While both the osmotic pressure and CD concentration affect the steady state flux in both permeation units, effects of sampling time and the MWCO of the mounted membrane were only observed in the Franz diffusion cells. The osmotic effect was negligible in the Slide-A-Lyzer™ MINI Dialysis units. The modified permeation technique using Slide-A-Lyzer™ MINI Dialysis units was then used to determine the cac of natural CDs in water. The cac of αCD, ßCD and γCD was 1.19±0.17, 0.69±0.05 and 0.93±0.04% (w/v), respectively. The results indicated that the cac values depended on their intrinsic solubility. Moreover, the cac value of γCD in aqueous hydrocortisone/γCD inclusion complex solution was identical to the γCD cac value determined in pure water.

Self-Assembly of Cyclodextrins and Their Complexes in Aqueous Solutions.

Cyclodextrins (CDs) are enabling pharmaceutical excipients that can be found in numerous pharmaceutical products worldwide. Because of their favorable toxicologic profiles, CDs are often used in toxicologic and phase I assessments of new drug candidates. However, at relatively high concentrations, CDs can spontaneously self-assemble to form visible microparticles in aqueous mediums and formation of such visible particles may cause product rejections. Formation of subvisible CD aggregates are also known to affect analytical results during product development. How and why these CD aggregates form is largely unknown, and factors contributing to their formation are still not elucidated. The physiochemical properties of CDs are very different from simple amphiphiles and lipophilic molecules that are known to self-assemble and form aggregates in aqueous solutions but very similar to those of linear oligosaccharides. In general, negligible amounts of aggregates are formed in pure CD solutions, but the aggregate formation is greatly enhanced on inclusion complex formation, and the extent of aggregation increases with increasing CD concentration. The diameter of the aggregates formed is frequently less than about 300 nm, but visible aggregates can also be formed under certain conditions.

A New Approach for Quantitative Determination of γ-Cyclodextrin in Aqueous Solutions: Application in Aggregate Determinations and Solubility in Hydrocortisone/γ-Cyclodextrin Inclusion Complex.

Fast and simple high-pressure liquid chromatographic (HPLC) method with charged aerosol detector (CAD) was developed for quantitation of γ-cyclodextrin (γCD) in aqueous solutions. The chromatographic system consisted of a C18 column (i.e., the stationary phase) and an aqueous mobile phase containing 7% (v/v) methanol. Calibration curve was obtained over the γCD concentration range of 0.005%-1% (w/v). The limit of detection and quantitation of γCD were 0.0001% and 0.0002% (w/v), respectively. Formation of γCD aggregates in aqueous solution and their critical aggregation concentration (cac) were determined by both conventional dynamic light scattering method and permeation method using HPLC-CAD for quantitative determination of γCD. The cac of γCD was determined to be 0.95% (w/v) and the amount of γCD self-aggregates increased with increasing γCD concentrations. Also, the developed HPLC-CAD method was used to determine the γCD phase-solubility profile in an aqueous hydrocortisone (HC)/γCD complexation medium. The maximum concentration of dissolved γCD and HC was determined to be 1.47% and 0.31% (w/v), respectively. The membrane permeation method was shown to be a reliable method for determination of metastable γCD aggregates. The HPLC-CAD method was successfully applied for quantitative determination of γCD in aqueous solutions during permeation and phase-solubility studies.

Sulfobutylether-ß-cyclodextrin/chitosan nano- and microparticles and their physicochemical characteristics.

Metastable polymer/cyclodextrin nano- and microparticles (NPs) were prepared from low molecular weight chitosan (CS), Mw about 10 kDa, and sulfobutylether ß-cyclodextrin (SBEßCD). CS is a cationic polysaccharide containing numerous protonated amino groups (pKa about 6.5). SBEßCD is a ß-cyclodextrin derivative with six to seven negatively charged sulfobutyl ether groups per cyclodextrin molecule. Ionotropic gelation technique was used to prepare the NPs. The NP matrix was composed of low molecular weight cationic CS polysaccharide cross-linked with polyvalent anions (SBEßCD). The diameter of the NPs ranged from 200 to almost 1,000 nm and was controlled by the CS:SBEßCD molar ratio during NP preparation. Hydrocortisone (HC) is a lipophilic drug with limited aqueous solubility (0.3mg/ml). HC displayed AL-type phase-solubility diagrams in aqueous solutions containing either SBEßCD or CS, although CS had negligible solubilizing effect. The ability of the NPs to encapsulate HC decreased with increasing CS concentration during preparation of the NPs even though the SBEßCD content of the NPs increased with increasing CS concentration. This decrease in HC encapsulation was related to the concentration; the ionic crosslinking provides better encapsulation at low initial SBEßCD and CS concentrations.