Effect of γ-cyclodextrin as a lyoprotectant for freeze-dried actinidin.

Actinidin (ATD) is a cysteine protease found in kiwifruit. It is used to tenderize meat and to enhance the digestion of proteins in the small intestine. However, ATD is unstable during freeze-drying, which alters its bioactivity. It is well known that sugars have the ability to protect proteins from the stress of freeze-drying. In this study, we investigated the protective effect of various saccharides on the stability of ATD during freeze-drying. The ATD activities of the samples containing γ-cyclodextrin (CyD) showed only a small decrease, and compared with trehalose and sucrose, γ-CyD was a more effective stabilizer for ATD. Secondary structural changes in freeze-dried ATD were observed by circular dichroism spectroscopy and compared with the changes in stabilized samples. There was a close relationship between the α-helix content and the stabilization. The sugars stabilized the protein by suppressing the changes in the α-helix. Fourier transform infrared spectroscopy measurement showed that the amide I band of ATD with γ-CyD was shifted to a lower wavenumber compared with other sugars. Therefore, stronger hydrogen bonds may be formed between ATD and γ-CyD than between ATD and other sugars. The suppression of changes in the protein secondary structure accompanying the formation of hydrogen bonding between the protein and the sugar also contributed to the protective effect of the sugars.

Effects of kaempferol on the mechanisms of drug resistance in the human glioblastoma cell line T98G.

Food contains components that may either increase or decrease the bioavailability of a drug. In particular, it is known that grapefruit juice and St. John's Wort induce drug interactions via an effect on the drug-metabolizing enzyme cytochrome P450 (CYP). However, interactions with membrane transporters, such as P-glycoprotein and multidrug resistance-related protein (MRP), may also influence drug bioavailability. The objective of the present study was to investigate the effects of kaempferol, a flavonoid present in food, on the cytotoxicity of anticancer drugs and the mechanisms of drug resistance in the human glioblastoma cell line T98G. Acute exposure to kaempferol inhibited the efflux of calcein, a substrate of MRP; however, chronic exposure caused no apparent effect on calcein efflux. The cytotoxicity of doxorubicin was not influenced by chronic exposure of cells to kaempferol, although that of cisplatin was significantly reduced. Multidrug resistance is often associated with increased levels of MRP1, glutathione S-transferase (GST) and activity by chronic exposure to kaempferol, although MRP2 protein levels are decreased. Accordingly, we hypothesized that the cytotoxicity of anticancer drugs that conjugate with glutathione and the substrate of MRPs may be influenced by long-term intake of drugs such as kaempferol, which are substrates of MRPs and GST.

Effectiveness of mechanochemical treatment with cyclodextrins on increasing solubility of glimepiride.

We investigated the enhancement of the solubility of glimepiride (GLM), a poorly water soluble anti-diabetes drug, by cogrinding it with various cyclodextrins (CDs) using a ball mill. The phase solubility profiles of GLM with beta-CD and its derivatives were classified as A(L)-type, indicating the formation of a 1 : 1 stoichiometric water-soluble complex. When GLM crystals were coground with beta-CD using a ball mill for 48 h, the aqueous solubility of GLM increased to approximately 250 microg/mL. The powder X-ray diffraction pattern showed that the peak intensity of crystalline GLM decreased after cogrinding. Endothermic peaks of around 208 degrees C, which were assigned to the fusion of GLM crystals, disappeared in the DSC measurement of the ground mixture. After cogrinding, two sharp peaks assigned to sulfonylurea and benzoyl carbonyl stretching bands varied to broaden the peak to around 1700 cm(-1) in the C=O stretching region. These results suggested the formation of a complex between GLM and beta-CD during cogrinding.

Improvement of solubility of C70 by complexation with cyclomaltononaose (delta-cyclodextrin).

We investigated the solubilizing effects of cyclomaltononaose (delta-CD), a cyclic oligosaccharide composed of nine alpha-1,4-linked D-glucose units, on C70 by using the ball-milling method based on a solid-solid mechanochemical reaction. The complex between C70 and delta-CD was characterized by UV-VIS spectrometry and fast atom bombardment mass spectrometry (FAB-MS). Coloration of the C70/delta-CD system was red-brown in aqueous solution, and the UV-VIS spectrum was in agreement with that of C70 in hexane solution. The FAB-MS spectrum of the C70/delta-CD system showed a negative ion peak corresponding to the molecular weight of a complex between two delta-CD and one C70. These findings suggest that the solubilization of C70 in water was due to complex formation of C70 with delta-CD, and the stoichiometric ratio of this complex was 1: 2 (C70: delta-CD).