Cholesterol-Lowering Effect of Octaarginine-Appended β-Cyclodextrin in Npc1-Trap-CHO Cells.
Niemann-Pick disease type C (NPC) is an autosomal recessive lysosomal storage disorder, which is an inherited disease characterized by the accumulation of unesterified cholesterol in endolysosomes. Recently, 2-hydroxypropyl-β-cyclodextrin (HP-β-CyD) has been used for the treatment of NPC, and ameliorated a hepatosplenomegaly in the patients. However, to obtain the treatment efficacy, a high dose of HP-β-CyD was necessary. Therefore, the decrease in dose by using active intracellular delivery system of β-CyD to NPC cells is expected. In this study, to efficiently deliver β-CyD to NPC-like cells, we newly synthesized octaarginine (R8)-appended β-CyD with a spacer of γ-aminobutyric acid (R8-β-CyD) and evaluated its cytotoxicity, intracellular distribution, endocytosis pathway and cholesterol-lowering effect in Npc1-trap-Chinese hamster ovary (CHO) cells, cholesterol-accumulated cells through the impairment of NPC1 function. R8-β-CyD did not show cytotoxicity in the cells. In addition, Alexa568-labeled R8-β-CyD was actively internalized into Npc1-trap-CHO cells, possibly through micropinocytosis. Notably, R8-β-CyD significantly decreased intracellular cholesterol content compared with HP-β-CyD. These results suggest that R8-β-CyD may be a promising therapeutic agent for ameliorating cholesterol accumulation in NPC.
Graduate School of Pharmaceutical Sciences, Kumamoto University.
Keiichi
K
Motoyama
Rena
R
Nishiyama
Yuki
Y
Maeda
Taishi
T
Higashi
Yoshimasa
Y
Kawaguchi
Shiroh
S
Futaki
Yoichi
Y
Ishitsuka
Yuki
Y
Kondo
Tetsumi
T
Irie
Takumi
T
Era
Hidetoshi
H
Arima
D000818:Animals; D016466:CHO Cells; D002784:Cholesterol / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D004705:Endocytosis; D052556:Niemann-Pick Disease, Type C; D009842:Oligopeptides / Q000737:chemistry / Q000494:pharmacology*; D047392:beta-Cyclodextrins / Q000737:chemistry / Q000494:pharmacology*; D005680:gamma-Aminobutyric Acid / Q000737:chemistry / Q000494:pharmacology*
D009842:Oligopeptides; D047392:beta-Cyclodextrins; C448619:octaarginine; D005680:gamma-Aminobutyric Acid; D002784:Cholesterol; C031215:betadex