Capsule shell material impacts the in vitro disintegration and dissolution behaviour of a green tea extract.

PURPOSE: In vitro disintegration and dissolution are routine methods used to assess the performance and quality of oral dosage forms. The purpose of the current work was to determine the potential for interaction between capsule shell material and a green tea extract and the impact it can have on the release. METHODS: A green tea extract was formulated into simple powder-in-capsule formulations of which the capsule shell material was either of gelatin or HPMC origin. The disintegration times were determined together with the dissolution profiles in compendial and biorelevant media. RESULTS: All formulations disintegrated within 30 min, meeting the USP criteria for botanical formulations. An immediate release dissolution profile was achieved for gelatin capsules in all media but not for the specified HPMC formulations. Dissolution release was especially impaired for HPMCgell at pH 1.2 and for both HPMC formulations in FeSSIF media suggesting the potential for food interactions. CONCLUSIONS: The delayed release from studied HPMC capsule materials is likely attributed to an interaction between the catechins, the major constituents of the green tea extract, and the capsule shell material. An assessment of in vitro dissolution is recommended prior to the release of a dietary supplement or clinical trial investigational product to ensure efficacy.

Caki-1 cells as a model system for the interaction of renally secreted drugs with OCT3.

BACKGROUND/AIMS: Organic cation transporters (OCT) in the proximal tubules (PTs) participate in the renal secretion of several therapeutic agents. The exact role of OCT3 in renal secretion remains undetermined, partially due to the lack of an appropriate in vitro model system. The current work introduces the PT representative cell line, Caki-1, as a model system for studying the involvement of OCT3 in renal secretion. METHODS: Caki-1 cells were characterized for OCT3 expression via real-time RT-PCR and immunocytochemical staining techniques. Uptake kinetics of OCT3 in Caki-1 cells was determined using prototypical substrates and inhibitors. Inhibition of OCT3-mediated uptake via several renally secreted drugs and those specifically of quaternary ammonium structure were determined. RESULTS: OCT3 expression was confirmed at the gene level and subcellular localization to the basolateral membrane (BLM) was illustrated for the first time. Caki-1 cells exhibited trademark kinetics of OCT3 and interacted with all therapeutic agents tested with varying affinities. The apparent IC(50 )values for cimetidine and trimethoprim were pharmacologically relevant. CONCLUSION: Confirmation for the usefulness of Caki-1 cells as a PT model system for investigations of OCT3 was obtained, a novel BLM localization of OCT3 was possible and relevant interactions between OCT3 and renally secreted drugs were shown.

Caki-1 cells represent an in vitro model system for studying the human proximal tubule epithelium.

BACKGROUND/AIMS: The human proximal tubule (PT) epithelium is distinguished from other nephron segments via several unique characteristics. Studies assessing PT epithelium increasingly employ cell lines, bypassing the complexity of primary cell cultures. However, few human model systems exist for studying PT cells in vitro. The current work involves an intensive characterization of Caki-1 cells, a commercially available human renal cell line. METHODS: Caki-1 cells were validated as a representative model system for PT cell research via morphological, physiological and biochemical investigations including light and transmission electron microscopy, transepithelial electrical resistance (TER) measurements and the detection of PT markers. RESULTS: Morphologically, these cells form a polarized monolayer with apical located microvilli and multiple mitochondria per cell. Low TER ranging from 2 to 28 Omega cm(2) was determined for Caki-1 cells, characteristic of the 'leaky' PT epithelium in vivo. Expression of the PT markers: NHE3, GGT, DPP IV, APM and AP were present in Caki-1 cells. Two epithelial markers, E-cadherin and Na(+)/K(+)-ATPase, were additionally observed. CONCLUSION: The current work is a concise summary which confirms that Caki-1 cells represent well-differentiated polarized PT cells in vitro, regardless of its cancerous origin and multiple passaging. They prove to be a significant contribution to the field of PT research.

OCTN2-mediated carnitine uptake in a newly discovered human proximal tubule cell line (Caki-1).

The proximal tubular reabsorption of carnitine in the human kidney is significant because more than 95% of the carnitine filtered in the kidney is reabsorbed by the proximal tubules therefore maintaining the homeostatic balance of carnitine in the body. Objectives of this study include the characterization of OCTN2 function in the Caki-1 cell line and the potential interactions of carnitine uptake with renally secreted drugs, including drugs of quaternary ammonium structure. Caki-1 cells were additionally characterized to be of proximal tubule nature, and an apical membrane expression pattern of OCTN2 in Caki-1 cells was discovered. Uptake studies with radiolabeled L-carnitine in Caki-1 cells revealed a Na+- and temperature-dependent carrier-mediated process (K(m) = 15.90 microM) which was unaffected by pH in a range from 6.5 to 8.5. All drugs tested were able to inhibit the carnitine uptake process to different degrees. The quaternary ammonium compounds ciclotropium bromide and ipratropium bromide were strong inhibitors with IC(50) values of 30 microM and 95 microM, respectively. The observed kinetics, immunohistolocalization, and inhibition studies indicate that the high-affinity uptake of carnitine in the Caki-1 cell line is most likely mediated by OCTN2. The interaction of drugs at the renal level with OCTN2 indicates a possible pathway for the final step of cationic secretion into the urine.

Different dissolution media lead to different crystal structures of talinolol with impact on its dissolution and solubility.

During the performance of dissolution tests with immediate and controlled-release talinolol tablets it was detected that the type of the buffer used as dissolution medium had a strong influence on the solubility and the dissolution behavior of the drug. It was proven that talinolol appeared in different crystal structures with strongly differing solubilities when pure water, acetate, or phosphate buffers were employed as dissolution media. The resulting crystal structures were characterized by means of light microscopy, differential scanning calorimetry, and X-ray powder diffraction. All methods were adjuvant to detect changes in talinolol crystal structures. The different solubility and dissolution properties of the talinolol salts or modifications may be viewed as a source for its incomplete and variable bioavailability. Furthermore, the food effect, described in the literature, that leads to a decrease in talinolol absorption, could be due to changes in the composition of gastrointestinal fluids leading to different talinolol crystal structures. Furthermore, it was detected that the addition of sodium chloride increases talinolol solubility and accelerates its dissolution from controlled-release tablets at concentrations between 0% and 1.25%, while an addition of sodium dodecylsulfate (SDS) as surfactant only had a solubility-improving effect at concentrations > 0.75%. At lower concentrations SDS decreased the solubility of talinolol and notably decelerated its release from controlled-release tablets.