A Fluorescent sp2-iminosugar with pharmacological chaperone activity for gaucher disease: synthesis and intracellular distribution studies.
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-(5N,6S)-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-(5N,6S)-[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.
Division of Child Neurology, Institute of Neurological Sciences, Faculty of Medicine, Tottori University, Nishi-cho, Yonago, 683-8504, Japan.
Zhuo
Z
Luan
Katsumi
K
Higaki
Matilde
M
Aguilar-Moncayo
Linjing
L
Li
Haruaki
H
Ninomiya
Eiji
E
Nanba
Kousaku
K
Ohno
M Isabel
MI
García-Moreno
Carmen
C
Ortiz Mellet
José M
JM
García Fernández
Yoshiyuki
Y
Suzuki
D017485:1-Deoxynojirimycin / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000493:pharmacokinetics; D000818:Animals; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D004795:Enzyme Stability; D005347:Fibroblasts / Q000378:metabolism; D005453:Fluorescence; D005776:Gaucher Disease / Q000201:enzymology* / Q000473:pathology; D005962:Glucosylceramidase / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D006358:Hot Temperature; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D042541:Intracellular Space / Q000378:metabolism; D008247:Lysosomes / Q000378:metabolism; D018613:Microscopy, Confocal; D008956:Models, Chemical; D018832:Molecular Chaperones / Q000138:chemical synthesis / Q000493:pharmacokinetics / Q000494:pharmacology; D015394:Molecular Structure; D009154:Mutation
C557857:6-thio-(5N,6S)-(4-(N'-dansylamino)butyliminomethylidene)nojirimycin; D018832:Molecular Chaperones; D017485:1-Deoxynojirimycin; D005962:Glucosylceramidase; C013626:nojirimycin