3-epi-fagomine and fagomine

A library of sp(2)-iminosugar conjugates derived from the piperidine iminosugar d-fagomine and the enantiomeric pyrrolidine iminosugars DAB and LAB has been generated in only two steps involving direct coupling of the fully unprotected polyhydroxylated heterocycles with isothiocyanates, to give monocyclic thiourea adducts, and further intramolecular nucleophilic displacement of the δ-located primary hydroxyl group by the thiocarbonyl sulphur atom, affording bicyclic isothioureas. These transformations led to a dramatic shift in the inhibitory selectivity from α- to β-glucosidases, with inhibition potencies that depended strongly on the nature of the aglycone-type moiety in the conjugates. Some of the new derivatives behaved as potent inhibitors of human β-glucocerebrosidase (GCase), the lysosomal enzyme whose dysfunction is responsible for Gaucher disease. Moreover, GCase inhibition was 10-fold weaker at pH 5 as compared to pH 7, which is generally considered as a good property for pharmacological chaperones. Surprisingly, most of the compounds strongly inhibited GCase in wild type fibroblasts at rather low concentrations, showing an unfavourable chaperone/inhibitor balance on disease-associated GCase mutants in cellulo. A structure-activity relationship analysis points to the need for keeping a contiguous triol system in the glycone moiety of the conjugates to elicit a chaperone effect. In any case, the results reported here represent a proof of concept of the utmost importance of implementing diversity-oriented strategies for the identification and optimization of potent and specific glycosidase inhibitors and chaperones.

Topics: Enzyme Inhibitors; Fibroblasts; Gaucher Disease; Glycoside Hydrolases; Humans; Imino Pyranoses; Imino Sugars; Structure-Activity Relationship

We report the structure-activity relationship of a series of D-, and L-isofagomine and fagomine isomers as glycosidase inhibitors. Our study revealed that a positive charge at the anomeric position of d-isofagomines enhanced the potency toward β-glycosidases, while the epimerization at the C3 OH group drastically reduced their inhibitory potency by over three orders of magnitude. Furthermore, d-3,4-di-epi-isofagomine abolished their inhibition activities against all enzymes. L-Isofagomine was also a fairly potent inhibitor of human β-glucocerebrosidase, with an IC₅₀ value of 8.7 μM. A molecular docking study revealed that the positions and orientations of the piperidine ring of D-3-epi-isofagomine in the binding site was similar to that of D-isofagomine, while D-3-epi-isofagomine missed the hydrogen bond interactions between Asp127 and the 3-OH group and between Trp179 and the 3-OH group. Furthermore, the top 10 docking models ranked by IFDscore suggested that D-3,4-di-epi-isofagomine can not bind to β-glucocerebrosidase at a stable interaction mode. These results provide an insight into the structural requirements of isofagomine isomers for developing a new type of pharmacological chaperone for Gaucher disease.

Topics: Animals; Binding Sites; Cattle; Computer Simulation; Enzyme Inhibitors; Glucosylceramidase; Humans; Hydrogen Bonding; Imino Pyranoses; Isomerism; Rats; Structure-Activity Relationship

50% aqueous MeOH extracts from the leaves and roots of Xanthocercis zambesiaca (Leguminosae) were subjected to various ion-exchange column chromatographic steps to give fagomine (1), 3-epi-fagomine (2), 3,4-di-epi-fagomine (3), 3-O-beta-D-glucopyranosylfagomine (4), and 4-O-beta-D-glucopyranosylfagomine (5). Their structures were determined by spectroscopic analyses, particularly by extensive 1D and 2D NMR studies. Compounds 3 and 4 are new natural products. Compound 1 is a good inhibitor of isomaltase and certain alpha- and beta-galactosidases. Whereas 2 is a more potent inhibitor of isomaltase and beta-galactosidases than 1, it does not inhibit alpha-galactosidase. Compounds 3-5 exhibited no significant inhibition against the glycosidases used.

Topics: alpha-Galactosidase; Animals; beta-Galactosidase; beta-Glucosidase; Cattle; Enzyme Inhibitors; Glucosidases; Glucosides; Hydrolysis; Imino Pyranoses; Liver; Magnetic Resonance Spectroscopy; Piperidines; Plants, Medicinal; Rats; Seeds