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A Fluorescent sp 2 ‐Iminosugar With Pharmacological Chaperone Activity for Gaucher Disease: Synthesis and Intracellular Distribution Studies
Author(s) -
Luan Zhuo,
Higaki Katsumi,
AguilarMoncayo Matilde,
Li Linjing,
Ninomiya Haruaki,
Nanba Eiji,
Ohno Kousaku,
GarcíaMoreno M. Isabel,
Ortiz Mellet Carmen,
García Fernández José M.,
Suzuki Yoshiyuki
Publication year - 2010
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201000323
Subject(s) - chaperone (clinical) , iminosugar , chemical chaperone , intracellular , lysosome , internalization , glucocerebrosidase , chemistry , biochemistry , microbiology and biotechnology , biology , endoplasmic reticulum , enzyme , medicine , unfolded protein response , cell , pathology
Gaucher disease (GD) is the most prevalent lysosomal‐storage disorder, it is caused by mutations of acid β‐glucosidase (β‐glucocerebrosidase; β‐Glu). Recently, we found that bicyclic nojirimycin (NJ) derivatives of the sp 2 ‐iminosugar type, including the 6‐thio‐ N′ ‐octyl‐(5 N ,6 S )‐octyliminomethylidene derivative (6S‐NOI‐NJ), behaved as very selective competitive inhibitors of the lysosomal β‐Glu and exhibited remarkable chaperone activities for several GD mutations. To obtain information about the cellular uptake pathway and intracellular distribution of this family of chaperones, we have synthesized a fluorescent analogue that maintains the fused piperidine–thiazolidine bicyclic skeleton and incorporates a dansyl group in the N′ ‐substituent, namely 6‐thio‐(5 N ,6 S )‐[4‐( N′ ‐dansylamino)butyliminomethylidene]nojirimycin (6S‐NDI‐NJ). This structural modification does not significantly modify the biological activity of the glycomimetic as a chemical chaperone. Our study showed that 6S‐NDI‐NJ is mainly located in lysosome‐related organelles in both normal and GD fibroblasts, and the fluorescent intensity of 6S‐NDI‐NJ in the lysosome is related to the β‐Glu concentration level. 6S‐NDI‐NJ also can enter cultured neuronal cells and act as a chaperone. Competitive inhibition studies of 6S‐NDI‐NJ uptake in fibroblasts showed that high concentrations of D ‐glucose have no effect on chaperone internalization, suggesting that it enters the cells through glucose‐transporter‐independent mechanisms.