A Fluorescent sp(2)-Iminosugar With Pharmacological Chaperone Activity for Gaucher Disease: Synthesis and Intracellular Distribution Studies

作者：Luan Zhuo; Higaki Katsumi^*; Aguilar Moncayo Matilde; Li Linjing; Ninomiya Haruaki; Nanba Eiji; Ohno Kousaku; Isabel Garcia Moreno M; Ortiz Mellet Carmen; Garcia Fernandez Jose M; Suzuki Yoshiyuki

来源：ChemBioChem, 2010, 11(17): 2453-2464.

DOI：10.1002/cbic.201000323

• chaperones
• fluorescent probes
• Gaucher disease
• glucosidases
• iminosugars

Gaucher disease (GD) is the most prevalent lysosomal-storage disorder, it is caused by mutations of acid beta-glucosidase (beta-glucocerebrosidase; beta-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 beta-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)butyliminomethyli-dene] 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 beta-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.

• 出版日期2010-11-22