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Herman S Overkleeft - One of the best experts on this subject based on the ideXlab platform.
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in vivo inactivation of glycosidases by conduritol b epoxide and cyclophellitol as revealed by activity based protein profiling
FEBS Journal, 2019Co-Authors: Chilin Kuo, Wouter W Kallemeijn, Lindsey T Lelieveld, Mina Mirzaian, Iris Zoutendijk, Ayelet Vardi, Anthony H Futerman, Annemarie H Meijer, Herman P Spaink, Herman S OverkleeftAbstract:Glucocerebrosidase (GBA) is a lysosomal β-glucosidase degrading glucosylceramide. Its deficiency causes Gaucher disease (GD), a common lysosomal storage disorder. Carrying a genetic abnormality in GBA constitutes at present the largest genetic risk factor for Parkinson's disease (PD). Conduritol B epoxide (CBE), a mechanism-based irreversible inhibitor of GBA, is used to generate cell and animal models for investigations on GD and PD. However, CBE may have additional glycosidase targets besides GBA. Here, we present the first in vivo target engagement study for CBE, employing a suite of activity-based probes to visualize catalytic pocket occupancy of candidate off-target glycosidases. Only at significantly higher CBE concentrations, non-lysosomal glucosylceramidase (GBA2) and lysosomal α-glucosidase were identified as major off-targets in cells and zebrafish larvae. A tight, but acceptable window for selective inhibition of GBA in the brain of mice was observed. On the other hand, cyclophellitol, a closer glucose mimic, was found to inactivate with equal affinity GBA and GBA2 and therefore is not suitable to generate genuine GD-like models. This article is protected by copyright. All rights reserved.
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distinguishing the differences in β Glycosylceramidase folds dynamics and actions informs therapeutic uses
Journal of Lipid Research, 2018Co-Authors: Fredj Ben Bdira, Marta Artola, Herman S Overkleeft, Marcellus Ubbink, Johannes M F G AertsAbstract:Glycosyl hydrolases (GHs) are carbohydrate-active enzymes that hydrolyze a specific β-glycosidic bond in glycoconjugate substrates; β-glucosidases degrade glucosylceramide, a ubiquitous glycosphingolipid. GHs are grouped into structurally similar families that themselves can be grouped into clans. GH1, GH5, and GH30 glycosidases belong to clan A hydrolases with a catalytic (β/α)8 TIM barrel domain, whereas GH116 belongs to clan O with a catalytic (α/α)6 domain. In humans, GH abnormalities underlie metabolic diseases. The lysosomal enzyme glucocerebrosidase (family GH30), deficient in Gaucher disease and implicated in Parkinson disease etiology, and the cytosol-facing membrane-bound glucosylceramidase (family GH116) remove the terminal glucose from the ceramide lipid moiety. Here, we compare enzyme differences in fold, action, dynamics, and catalytic domain stabilization by binding site occupancy. We also explore other glycosidases with reported Glycosylceramidase activity, including human cytosolic β-glucosidase, intestinal lactase-phlorizin hydrolase, and lysosomal galactosylceramidase. Last, we describe the successful translation of research to practice: recombinant glycosidases and glucosylceramide metabolism modulators are approved drug products (enzyme replacement therapies). Activity-based probes now facilitate the diagnosis of enzyme deficiency and screening for compounds that interact with the catalytic pocket of glycosidases. Future research may deepen the understanding of the functional variety of these enzymes and their therapeutic potential.
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the development of an aza c glycoside library based on a tandem staudinger aza wittig ugi three component reaction
European Journal of Organic Chemistry, 2012Co-Authors: Johannes M F G Aerts, Richard J B H N Van Den Berg, Tom Wennekes, Kimberley M Bonger, Katrin Vogel, Anneke Strijland, Wilma E Donkerkoopman, Gijsbert A Van Der Marel, Herman S OverkleeftAbstract:We report the tandem Staudinger/aza-Wittig/Ugi three-component reaction mediated synthesis of a 64-member compound library of aza-C-glycosides. The library is composed of four pyrrolidine and three piperidine scaffolds, onto which a number of functional groups is grafted to form seven sublibraries. Variation in the library is achieved by transformation of two pentoses and a hexose into the corresponding 4-azidopentanal and 5-azidohexanal derivatives as precursors for the Staudinger/aza-Wittig process. Further variation is achieved by using different isocyanides as well as protective- and functional-group manipulations on the fully protected Ugi-3CR intermediates. Preliminary biological evaluation of the compound library revealed several low micromolar inhibitors of human acid glucosylceramidase
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generation of specific deoxynojirimycin type inhibitors of the non lysosomal glucosylceramidase
Journal of Biological Chemistry, 1998Co-Authors: Herman S Overkleeft, G H Renkema, J Neele, P Vianello, I O Hung, A Strijland, A M Van Der Burg, G J Koomen, U K Pandit, Johannes M F G AertsAbstract:The existence of a non-lysosomal glucosylceramidase in human cells has been documented (van Weely, S., Brandsma, M., Strijland, A., Tager, J. M., and Aerts, J. M. F. G. (1993) Biochim. Biophys. Acta 1181, 55-62). Hypothetically, the activity of this enzyme, which is localized near the cell surface, may influence ceramide-mediated signaling processes. To obtain insight in the physiological importance of the non-lysosomal glucosylceramidase, the availability of specific inhibitors would be helpful. Here we report on the generation of hydrophobic deoxynojirimycin (DNM) derivatives that potently inhibit the enzyme. The inhibitors were designed on the basis of the known features of the non-lysosomal glucosylceramidase and consist of a DNM moiety, an N-alkyl spacer, and a large hydrophobic group that promotes insertion in membranes. In particular, N-(5-adamantane-1-yl-methoxy)pentyl)-DNM is a very powerful inhibitor of the non-lysosomal glucosylceramidase at nanomolar concentrations. At such concentrations, the lysosomal glucocerebrosidase and alpha-glucosidase, the glucosylceramide synthase, and the N-linked glycan-trimming alpha-glucosidases of the endoplasmic reticulum are not affected.
Johannes M F G Aerts - One of the best experts on this subject based on the ideXlab platform.
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distinguishing the differences in β Glycosylceramidase folds dynamics and actions informs therapeutic uses
Journal of Lipid Research, 2018Co-Authors: Fredj Ben Bdira, Marta Artola, Herman S Overkleeft, Marcellus Ubbink, Johannes M F G AertsAbstract:Glycosyl hydrolases (GHs) are carbohydrate-active enzymes that hydrolyze a specific β-glycosidic bond in glycoconjugate substrates; β-glucosidases degrade glucosylceramide, a ubiquitous glycosphingolipid. GHs are grouped into structurally similar families that themselves can be grouped into clans. GH1, GH5, and GH30 glycosidases belong to clan A hydrolases with a catalytic (β/α)8 TIM barrel domain, whereas GH116 belongs to clan O with a catalytic (α/α)6 domain. In humans, GH abnormalities underlie metabolic diseases. The lysosomal enzyme glucocerebrosidase (family GH30), deficient in Gaucher disease and implicated in Parkinson disease etiology, and the cytosol-facing membrane-bound glucosylceramidase (family GH116) remove the terminal glucose from the ceramide lipid moiety. Here, we compare enzyme differences in fold, action, dynamics, and catalytic domain stabilization by binding site occupancy. We also explore other glycosidases with reported Glycosylceramidase activity, including human cytosolic β-glucosidase, intestinal lactase-phlorizin hydrolase, and lysosomal galactosylceramidase. Last, we describe the successful translation of research to practice: recombinant glycosidases and glucosylceramide metabolism modulators are approved drug products (enzyme replacement therapies). Activity-based probes now facilitate the diagnosis of enzyme deficiency and screening for compounds that interact with the catalytic pocket of glycosidases. Future research may deepen the understanding of the functional variety of these enzymes and their therapeutic potential.
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the development of an aza c glycoside library based on a tandem staudinger aza wittig ugi three component reaction
European Journal of Organic Chemistry, 2012Co-Authors: Johannes M F G Aerts, Richard J B H N Van Den Berg, Tom Wennekes, Kimberley M Bonger, Katrin Vogel, Anneke Strijland, Wilma E Donkerkoopman, Gijsbert A Van Der Marel, Herman S OverkleeftAbstract:We report the tandem Staudinger/aza-Wittig/Ugi three-component reaction mediated synthesis of a 64-member compound library of aza-C-glycosides. The library is composed of four pyrrolidine and three piperidine scaffolds, onto which a number of functional groups is grafted to form seven sublibraries. Variation in the library is achieved by transformation of two pentoses and a hexose into the corresponding 4-azidopentanal and 5-azidohexanal derivatives as precursors for the Staudinger/aza-Wittig process. Further variation is achieved by using different isocyanides as well as protective- and functional-group manipulations on the fully protected Ugi-3CR intermediates. Preliminary biological evaluation of the compound library revealed several low micromolar inhibitors of human acid glucosylceramidase
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Iminosugar-based inhibitors of glucosylceramide synthase increase brain glycosphingolipids and survival in a mouse model of Sandhoff disease
PLOS ONE, 2011Co-Authors: Kathleen M Ashe, Jennifer B Nietupski, Mario A. Cabrera-salazar, Christopher G.f. Cooper, Dinesh S Bangari, Lingyun Li, Johannes M F G Aerts, Scott D Bercury, Diane P CopelandAbstract:The neuropathic glycosphingolipidoses are a subgroup of lysosomal storage disorders for which there are no effective therapies. A potential approach is substrate reduction therapy using inhibitors of glucosylceramide synthase (GCS) to decrease the synthesis of glucosylceramide and related glycosphingolipids that accumulate in the lysosomes. Genz-529468, a blood-brain barrier-permeant iminosugar-based GCS inhibitor, was used to evaluate this concept in a mouse model of Sandhoff disease, which accumulates the glycosphingolipid GM2 in the visceral organs and CNS. As expected, oral administration of the drug inhibited hepatic GM2 accumulation. Paradoxically, in the brain, treatment resulted in a slight increase in GM2 levels and a 20-fold increase in glucosylceramide levels. The increase in brain glucosylceramide levels might be due to concurrent inhibition of the non-lysosomal glucosylceramidase, Gba2. Similar results were observed with NB-DNJ, another iminosugar-based GCS inhibitor. Despite these unanticipated increases in glycosphingolipids in the CNS, treatment nevertheless delayed the loss of motor function and coordination and extended the lifespan of the Sandhoff mice. These results suggest that the CNS benefits observed in the Sandhoff mice might not necessarily be due to substrate reduction therapy but rather to off-target effects.
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generation of specific deoxynojirimycin type inhibitors of the non lysosomal glucosylceramidase
Journal of Biological Chemistry, 1998Co-Authors: Herman S Overkleeft, G H Renkema, J Neele, P Vianello, I O Hung, A Strijland, A M Van Der Burg, G J Koomen, U K Pandit, Johannes M F G AertsAbstract:The existence of a non-lysosomal glucosylceramidase in human cells has been documented (van Weely, S., Brandsma, M., Strijland, A., Tager, J. M., and Aerts, J. M. F. G. (1993) Biochim. Biophys. Acta 1181, 55-62). Hypothetically, the activity of this enzyme, which is localized near the cell surface, may influence ceramide-mediated signaling processes. To obtain insight in the physiological importance of the non-lysosomal glucosylceramidase, the availability of specific inhibitors would be helpful. Here we report on the generation of hydrophobic deoxynojirimycin (DNM) derivatives that potently inhibit the enzyme. The inhibitors were designed on the basis of the known features of the non-lysosomal glucosylceramidase and consist of a DNM moiety, an N-alkyl spacer, and a large hydrophobic group that promotes insertion in membranes. In particular, N-(5-adamantane-1-yl-methoxy)pentyl)-DNM is a very powerful inhibitor of the non-lysosomal glucosylceramidase at nanomolar concentrations. At such concentrations, the lysosomal glucocerebrosidase and alpha-glucosidase, the glucosylceramide synthase, and the N-linked glycan-trimming alpha-glucosidases of the endoplasmic reticulum are not affected.
Tom Wennekes - One of the best experts on this subject based on the ideXlab platform.
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a fluorescence polarization activity based protein profiling assay in the discovery of potent selective inhibitors for human nonlysosomal glucosylceramidase
Journal of the American Chemical Society, 2017Co-Authors: Daniel Lahav, Richard J B H N Van Den Berg, Adrianus M C H Van Den Nieuwendijk, Tom Wennekes, Amar B T Ghisaidoobe, Imogen Z Breen, Maria J Ferraz, Liang Wu, Paul P Geurink, Huib OvaaAbstract:Human nonlysosomal glucosylceramidase (GBA2) is one of several enzymes that controls levels of glycolipids and whose activity is linked to several human disease states. There is a major need to design or discover selective GBA2 inhibitors both as chemical tools and as potential therapeutic agents. Here, we describe the development of a fluorescence polarization activity-based protein profiling (FluoPol-ABPP) assay for the rapid identification, from a 350+ library of iminosugars, of GBA2 inhibitors. A focused library is generated based on leads from the FluoPol-ABPP screen and assessed on GBA2 selectivity offset against the other glucosylceramide metabolizing enzymes, glucosylceramide synthase (GCS), lysosomal glucosylceramidase (GBA), and the cytosolic retaining β-glucosidase, GBA3. Our work, yielding potent and selective GBA2 inhibitors, also provides a roadmap for the development of high-throughput assays for identifying retaining glycosidase inhibitors by FluoPol-ABPP on cell extracts containing recomb...
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the development of an aza c glycoside library based on a tandem staudinger aza wittig ugi three component reaction
European Journal of Organic Chemistry, 2012Co-Authors: Johannes M F G Aerts, Richard J B H N Van Den Berg, Tom Wennekes, Kimberley M Bonger, Katrin Vogel, Anneke Strijland, Wilma E Donkerkoopman, Gijsbert A Van Der Marel, Herman S OverkleeftAbstract:We report the tandem Staudinger/aza-Wittig/Ugi three-component reaction mediated synthesis of a 64-member compound library of aza-C-glycosides. The library is composed of four pyrrolidine and three piperidine scaffolds, onto which a number of functional groups is grafted to form seven sublibraries. Variation in the library is achieved by transformation of two pentoses and a hexose into the corresponding 4-azidopentanal and 5-azidohexanal derivatives as precursors for the Staudinger/aza-Wittig process. Further variation is achieved by using different isocyanides as well as protective- and functional-group manipulations on the fully protected Ugi-3CR intermediates. Preliminary biological evaluation of the compound library revealed several low micromolar inhibitors of human acid glucosylceramidase
Huib Ovaa - One of the best experts on this subject based on the ideXlab platform.
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a fluorescence polarization activity based protein profiling assay in the discovery of potent selective inhibitors for human nonlysosomal glucosylceramidase
Journal of the American Chemical Society, 2017Co-Authors: Daniel Lahav, Richard J B H N Van Den Berg, Adrianus M C H Van Den Nieuwendijk, Tom Wennekes, Amar B T Ghisaidoobe, Imogen Z Breen, Maria J Ferraz, Liang Wu, Paul P Geurink, Huib OvaaAbstract:Human nonlysosomal glucosylceramidase (GBA2) is one of several enzymes that controls levels of glycolipids and whose activity is linked to several human disease states. There is a major need to design or discover selective GBA2 inhibitors both as chemical tools and as potential therapeutic agents. Here, we describe the development of a fluorescence polarization activity-based protein profiling (FluoPol-ABPP) assay for the rapid identification, from a 350+ library of iminosugars, of GBA2 inhibitors. A focused library is generated based on leads from the FluoPol-ABPP screen and assessed on GBA2 selectivity offset against the other glucosylceramide metabolizing enzymes, glucosylceramide synthase (GCS), lysosomal glucosylceramidase (GBA), and the cytosolic retaining β-glucosidase, GBA3. Our work, yielding potent and selective GBA2 inhibitors, also provides a roadmap for the development of high-throughput assays for identifying retaining glycosidase inhibitors by FluoPol-ABPP on cell extracts containing recomb...
Richard J B H N Van Den Berg - One of the best experts on this subject based on the ideXlab platform.
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a fluorescence polarization activity based protein profiling assay in the discovery of potent selective inhibitors for human nonlysosomal glucosylceramidase
Journal of the American Chemical Society, 2017Co-Authors: Daniel Lahav, Richard J B H N Van Den Berg, Adrianus M C H Van Den Nieuwendijk, Tom Wennekes, Amar B T Ghisaidoobe, Imogen Z Breen, Maria J Ferraz, Liang Wu, Paul P Geurink, Huib OvaaAbstract:Human nonlysosomal glucosylceramidase (GBA2) is one of several enzymes that controls levels of glycolipids and whose activity is linked to several human disease states. There is a major need to design or discover selective GBA2 inhibitors both as chemical tools and as potential therapeutic agents. Here, we describe the development of a fluorescence polarization activity-based protein profiling (FluoPol-ABPP) assay for the rapid identification, from a 350+ library of iminosugars, of GBA2 inhibitors. A focused library is generated based on leads from the FluoPol-ABPP screen and assessed on GBA2 selectivity offset against the other glucosylceramide metabolizing enzymes, glucosylceramide synthase (GCS), lysosomal glucosylceramidase (GBA), and the cytosolic retaining β-glucosidase, GBA3. Our work, yielding potent and selective GBA2 inhibitors, also provides a roadmap for the development of high-throughput assays for identifying retaining glycosidase inhibitors by FluoPol-ABPP on cell extracts containing recomb...
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the development of an aza c glycoside library based on a tandem staudinger aza wittig ugi three component reaction
European Journal of Organic Chemistry, 2012Co-Authors: Johannes M F G Aerts, Richard J B H N Van Den Berg, Tom Wennekes, Kimberley M Bonger, Katrin Vogel, Anneke Strijland, Wilma E Donkerkoopman, Gijsbert A Van Der Marel, Herman S OverkleeftAbstract:We report the tandem Staudinger/aza-Wittig/Ugi three-component reaction mediated synthesis of a 64-member compound library of aza-C-glycosides. The library is composed of four pyrrolidine and three piperidine scaffolds, onto which a number of functional groups is grafted to form seven sublibraries. Variation in the library is achieved by transformation of two pentoses and a hexose into the corresponding 4-azidopentanal and 5-azidohexanal derivatives as precursors for the Staudinger/aza-Wittig process. Further variation is achieved by using different isocyanides as well as protective- and functional-group manipulations on the fully protected Ugi-3CR intermediates. Preliminary biological evaluation of the compound library revealed several low micromolar inhibitors of human acid glucosylceramidase