title: New insights into the pharmacological chaperone activity of c2-substituted glucoimidazoles for the treatment of Gaucher disease.
abstract: Mutations in acid β-glucocerebrosidase (GCase) lead to the accumulation of the sphingolipid glucosylceramide, thereby resulting in Gaucher disease (GD). Active-site-specific competitive GCase inhibitors are effective pharmacological chaperones (PCs) that act as folding agents for mutant GCase folding in the endoplasmic reticulum. In this study, we prepared a series of glucoimidazole C2-substituent derivatives, and evaluated their inhibition and PC properties with GCase. A cell-based assay with patient-derived lymphoblasts (N370S or L444P mutations) demonstrated that administration of these compounds can significantly increase GCase activity. Interestingly, the 3,3-dimethyl-N-phenyl-4-amide-1-butyl-substituted moderate inhibitor 11 had the greatest effect on activity: 2.1-fold increase in N370S lymphoblasts at 2.5 μM and 1.2-fold increase in L444P at 0.5 μM following a three-day incubation. Computer docking studies and a protease protection assay were used to elucidate the ligand-enzyme interactions responsible for the chaperone activity of 11. Western blot and immuno-fluorescence assays verified restoration of GCase trafficking to the lysosome. Together, these results indicate that 11 is a promising PC for GD treatment and provide direct evidence of the mechanism of GCase chaperoning.
authors.affiliation: State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Nankai University, 94 Weijin Road, Tianjin 300071, P.R.China.
authors.forename: Zhonghua
authors.initials: Z
authors.lastname: Li
authors.forename: Tiehai
authors.initials: T
authors.lastname: Li
authors.forename: Shaoxing
authors.initials: S
authors.lastname: Dai
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Xie
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Ma
authors.forename: Wei
authors.initials: W
authors.lastname: Zhao
authors.forename: Weimin
authors.initials: W
authors.lastname: Zhang
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.forename: Peng George
authors.initials: PG
authors.lastname: Wang
mesh_terms: D005347:Fibroblasts / Q000187:drug effects; D005776:Gaucher Disease / Q000188:drug therapy* / Q000201:enzymology / Q000235:genetics; D005962:Glucosylceramidase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007093:Imidazoles / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D008214:Lymphocytes / Q000187:drug effects / Q000201:enzymology; D018613:Microscopy, Confocal; D008958:Models, Molecular; D018832:Molecular Chaperones / Q000737:chemistry / Q000494:pharmacology*; D009154:Mutation; D017510:Protein Folding
chemical_list: D007093:Imidazoles; D018832:Molecular Chaperones; D005962:Glucosylceramidase; C090568:imiglucerase
keywords: GCases; Gaucher disease; biological activity; chaperone proteins; glucoimidazoles; lysosomal storage diseases