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Design,Synthesis and Structural Analysis of Glucocerebrosidase Imaging Agents |
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| Authors: | Rhianna J. Rowland Yurong Chen Dr. Imogen Breen Dr. Liang Wu Wendy A. Offen Dr. Thomas J. Beenakker Qin Su Adrianus M. C. H. van den Nieuwendijk Prof. Johannes M. F. G. Aerts Dr. Marta Artola Prof. Herman S. Overkleeft Prof. Gideon J. Davies |
| Affiliation: | 1. Department of Chemistry, York Structural Biology Laboratory (YSBL), University of York Heslington, York, YO10 5DD UK These authors contributed equally to this work.;2. Department of Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden University, Einsteinwegg 55, 2300 RA Leiden, Netherlands These authors contributed equally to this work.;3. Department of Chemistry, York Structural Biology Laboratory (YSBL), University of York Heslington, York, YO10 5DD UK;4. Department of Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden University, Einsteinwegg 55, 2300 RA Leiden, Netherlands;5. Department of Medicinal Biochemistry, Leiden Institute of Chemistry, Leiden University, Einsteinwegg 55, 2300 RA Leiden, Netherlands |
| Abstract: | Gaucher disease (GD) is a lysosomal storage disorder caused by inherited deficiencies in β-glucocerebrosidase (GBA). Current treatments require rapid disease diagnosis and a means of monitoring therapeutic efficacy, both of which may be supported by the use of GBA-targeting activity-based probes (ABPs). Here, we report the synthesis and structural analysis of a range of cyclophellitol epoxide and aziridine inhibitors and ABPs for GBA. We demonstrate their covalent mechanism-based mode of action and uncover binding of the new N-functionalised aziridines to the ligand binding cleft. These inhibitors became scaffolds for the development of ABPs; the O6-fluorescent tags of which bind in an allosteric site at the dimer interface. Considering GBA's preference for O6- and N-functionalised reagents, a bi-functional aziridine ABP was synthesized as a potentially more powerful imaging agent. Whilst this ABP binds to two unique active site clefts of GBA, no further benefit in potency was achieved over our first generation ABPs. Nevertheless, such ABPs should serve useful in the study of GBA in relation to GD and inform the design of future probes. |
| Keywords: | Activity-based probes Carbohydrates Cyclitols Fluorescent Probes β-Glucocerebrosidase Inhibitors Structural Biology |