RESUMEN
A new class of compounds inhibiting de-O-glycosylation of proteins has been identified. Highly substituted diaminocyclopentanes are impressively selective reversible non-transition state O-ß-N-acetyl-d-glucosaminidase (O-GlcNAcase) inhibitors. The ease of preparative access and remarkable biological activities provide highly viable leads for the development of anti-tau-phosphorylation agents with a view to eventually ameliorating Alzheimer's disease.
Asunto(s)
Enfermedad de Alzheimer , beta-N-Acetilhexosaminidasas , Humanos , Hexosaminidasas , GlicosilaciónRESUMEN
Selective covalent labelling of enzymes using small molecule probes has advanced the scopes of protein profiling. The covalent bond formation to a specific target is the key step of activity-based protein profiling (ABPP), a method which has become an indispensable tool for measuring enzyme activity in complex matrices. With respect to carbohydrate processing enzymes, strategies for ABPP so far involve labelling the active site of the enzyme, which results in permanent loss of activity. Here, we report in a proof of concept study the use of ligand-directed chemistry (LDC) for labelling glycoside hydrolases near - but not in - the active site. During the labelling process, the competitive inhibitor is cleaved from the probe, departs the active site and the enzyme maintains its catalytic activity. To this end, we designed a building block synthetic concept for small molecule probes containing iminosugar-based reversible inhibitors for labelling of two model ß-glucosidases. The results indicate that the LDC approach can be adaptable for covalent proximity labelling of glycoside hydrolases.
Asunto(s)
Carbohidratos , Glicósido Hidrolasas , Glicósido Hidrolasas/metabolismo , Prueba de Estudio Conceptual , LigandosRESUMEN
A short survey on selected ß-galactosidase inhibitors as potential pharmacological chaperones for GM1 -gangliosidosis and Morquio B associated mutants of human lysosomal ß-galactosidase is provided highlighting recent developments in this particular area of lysosomal storage disorders and orphan diseases.
Asunto(s)
Gangliosidosis , Gangliosidosis GM1 , Mucopolisacaridosis IV , beta-Galactosidasa/antagonistas & inhibidores , Gangliosidosis GM1/tratamiento farmacológico , Humanos , Lisosomas , Mucopolisacaridosis IV/tratamiento farmacológicoRESUMEN
A set of cyclopentanoid α-galactosidase ligands was prepared from a partially protected ω-eno-aldose via a reliable (2 + 3)-cycloaddition protocol with slightly modified conditions. The obtained N-benzylisoxazolidine ring was selectively opened and the configuration of the hydroxymethylgroup was inverted. Consecutive deprotection provided an aminocyclopentane, which was N-alkylated to furnish a set of potential α-galactosidase inhibitors. Their glycosidase inhibitory activities were screened with a panel of standard glycosidases of biological significance.
RESUMEN
The scope of a series of N-alkylated iminosugar based inhibitors in the d-gluco as well as d-xylo configuration towards their interaction with human lysosomal ß-glucocerebrosidase has been evaluated. A versatile synthetic toolbox has been developed for the synthesis of N-alkylated iminosugar scaffolds conjugated to a variety of terminal groups via a benzoic acid ester linker. The terminal groups such as nitrile, azide, alkyne, nonafluoro-tert-butyl and amino substituents enable follow-up chemistry as well as visualisation experiments. All compounds showed promising inhibitory properties as well as selectivities for ß-glucosidases, some exhibiting activities in the low nanomolar range for ß-glucocerebrosidase.
Asunto(s)
Glucosilceramidasa/metabolismo , Lisosomas/enzimología , Ácido Benzoico/metabolismo , Humanos , Cinética , Espectroscopía de Resonancia Magnética , Estructura MolecularRESUMEN
Glycosidase inhibitors have shown great potential as pharmacological chaperones for lysosomal storage diseases. In light of this, a series of new cyclopentanoid ß-galactosidase inhibitors were prepared and their inhibitory and pharmacological chaperoning activities determined and compared with those of lipophilic analogs of the potent ß-d-galactosidase inhibitor 4-epi-isofagomine. Structure-activity relationships were investigated by X-ray crystallography as well as by alterations in the cyclopentane moiety such as deoxygenation and replacement by fluorine of a "strategic" hydroxyl group. New compounds have revealed highly promising activities with a range of ß-galactosidase-compromised human cell lines and may serve as leads towards new pharmacological chaperones for GM1-gangliosidosis and Morquio B disease.
Asunto(s)
Ciclopentanos/farmacología , Galactosidasas/metabolismo , Iminopiranosas/farmacología , Lisosomas/enzimología , Chaperonas Moleculares/metabolismo , Cristalización , Ciclopentanos/síntesis química , Ciclopentanos/química , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Galactosidasas/antagonistas & inhibidores , Humanos , Iminopiranosas/síntesis química , Iminopiranosas/química , Ligandos , Lisosomas/efectos de los fármacos , Conformación Molecular , Proteínas Mutantes/metabolismoRESUMEN
ABSTRACT: Modified 1,5-dideoxy-1,5-imino-d-xylitol analogues with different substitution patterns involving position C-1 and/or the ring nitrogen were prepared, which were designed to serve as precursors for the preparation of iminoxylitol-based ligands and tools for the elucidation and modulation of human lysosomal ß-glucocerebrosidase. Biological evaluation of the synthesized glycomimetics with a series of glycoside hydrolases revealed that these substitution patterns elicit excellent ß-glucosidase selectivities.
RESUMEN
A series of N-alkyl derivatives of the D-galactosidase inhibitor 1,4-di-epi-validamine featuring lipophilic substituents at position C-5a was prepared and screened for their glycosidase inhibitory properties. Products turned out selective for ß-galactosidases as well as ß-glucosidases.
Asunto(s)
Proteínas Bacterianas/antagonistas & inhibidores , Inhibidores Enzimáticos/química , Proteínas Fúngicas/antagonistas & inhibidores , Inositol/análogos & derivados , beta-Galactosidasa/antagonistas & inhibidores , beta-Glucosidasa/antagonistas & inhibidores , Agrobacterium/química , Agrobacterium/enzimología , Animales , Proteínas Bacterianas/química , Conformación de Carbohidratos , Bovinos , Inhibidores Enzimáticos/síntesis química , Escherichia coli/química , Escherichia coli/enzimología , Proteínas Fúngicas/química , Interacciones Hidrofóbicas e Hidrofílicas , Inositol/síntesis química , Inositol/química , Cinética , Saccharomyces cerevisiae/química , Saccharomyces cerevisiae/enzimología , beta-Galactosidasa/química , beta-Glucosidasa/químicaRESUMEN
From a secondary hydroxyl group, by the simple sequence of oxidation, Wittig reaction of the obtained ulose with methoxymethylene triphenyl phosphorane, exposure of the resulting exocyclic enol ether to Selectfluor and subsequent reduction of the α-fluoro aldehyde thus obtained, tertiary fluoro substituents can be introduced into carbohydrate and carbohydrate-related scaffolds at a branching point now bearing a new hydroxymethyl group.