Protective effect of sulfobutyl ether β-cyclodextrin on DY-9760e-induced hemolysis in vitro.
Y. Nagase 1, M. Hirata 2, H. Arima 2, S. Tajiri 2, Y. Nishimoto 2, F. Hirayama 2, T. Irie 2, K. Uekama 2 *
1Tokyo Pharmaceutical Research Center, Pharmaceutical Technology Research Laboratories, Daiichi Pharmaceutical Company, 1-16-13, Kitakasai, Edogawa-ku, Tokyo 134-8630, Japan
2Faculty of Pharmaceutical Sciences, Kumamoto University, 5-1 Oe-honmachi, Kumamoto 862-0973, Japan
Abstract
he hemolytic behavior of a novel cytoprotective agent, DY-9760e (3-[2-[4-(3-chloro-2-methylphenyl)-1-piperazinyl]ethyl]-5,6-dimethoxy-1-(4-imidazolylmethyl)-1H-indazole dihydrochloride 3.5 hydrate) was investigated using rabbit erythrocytes. Further, the effects of water-soluble cyclodextrin derivatives, such as 2-hydroxypropyl-β-cyclodextrin (HP-β-CyD) and sulfobutyl ether of β-cyclodextrin (SBE-β-CyD), on the hemolytic activity of DY-9760e were studied. DY-9760e induced hemolysis at concentrations >0.2-0.3 mM in phosphate buffered saline (PBS) of pH 4.0 and 6.0, where DY-9760e is predominantly in dicationic and monocationic forms, respectively. The hemolytic activity of the monocationic DY-9760e was higher than that of the dicationic species, and the hemolysis at pH 4.0 involved the formation of methemoglobin. DY9760e induced the morphological change of erythrocytes towards membrane invagination at both pH 4.0 and 6.0. SBE7-β-CyD significantly suppressed the DY-9760e-induced hemolysis and morphological change at both pH 4.0 and 6.0, as well as the formation of methemoglobin at pH 4.0. On the other hand, HP-β-CyD suppressed only the hemolysis, but neither the morphological change nor the formation of methemoglobin. In addition, the inhibitory effect of SBE7-β-CyD on the hemolysis was greater than that of HP-β-CyD. The superior inhibitory effect of SBE7-β-CyD on the DY-9760-induced hemolysis, the morphological change, and the formation of methemoglobin may be attributable to the formation of a stable inclusion complex with DY-9760e and to the weaker hemolytic activity of SBE7-CyD than HP-β-CyD. These results suggest potential use of SBE7-β-CyD as a parenteral carrier for DY-9760e. Wiley-Liss, Inc. and the American Pharmaceutical Association J Pharm Sci 91:2382-2389, 2002