Cited by 105Lysosomal Storage Disorders ResearchGlycogen Storage Diseases and MyoclonusCarbohydrate Chemistry and Synthesis

High-Throughput Screening for Human Lysosomal β-N-Acetyl Hexosaminidase Inhibitors Acting as Pharmacological Chaperones

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Tropak, Michael B., Blanchard, Jan E., Withers, Stephen G., Brown, Eric D., Mahuran, Don·2007·Chemistry & Biology

Abstract

The adult forms of Tay-Sachs and Sandhoff diseases result when the activity of beta-hexosaminidase A (Hex) falls below approximately 10% of normal due to decreased transport of the destabilized mutant enzyme to the lysosome. Carbohydrate-based competitive inhibitors of Hex act as pharmacological chaperones (PC) in patient cells, facilitating exit of the enzyme from the endoplasmic reticulum, thereby increasing the mutant Hex protein and activity levels in the lysosome 3- to 6-fold. To identify drug-like PC candidates, we developed a fluorescence-based real-time enzyme assay and screened the Maybridge library of 50,000 compounds for inhibitors of purified Hex. Three structurally distinct micromolar competitive inhibitors, a bisnaphthalimide, nitro-indan-1-one, and pyrrolo[3,4-d]pyridazin-1-one were identified that specifically increased lysosomal Hex protein and activity levels in patient fibroblasts. These results validate screening for inhibitory compounds as an approach to identifying PCs.

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Crossref · 10.1016/j.chembiol.2006.12.006

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Authors
Tropak, M. B., Blanchard, J. E., Withers, S. G., Brown, E. D., & Mahuran, D.
Affiliations
  • Hospital for Sick Children (CA)
  • McMaster University (CA)
  • University of British Columbia (CA)
  • University of Toronto (CA)
Corresponding author
Mahuran, Don
Venue
Chemistry & Biology
Publisher
Elsevier BV
Published
2007-02
Volume
14
Issue
2
Pages
153-164
ISSN
1074-5521 (ISSN-L)
OpenAlex
W2160928326
Cited by
105
References
58
Citations by year
2025: 3 · 2024: 2 · 2023: 4 · 2022: 5 · 2021: 4 · 2020: 3 · 2019: 4 · 2018: 6 · 2017: 4 · 2016: 4 · 2015: 2 · 2014: 10 · 2013: 6 · 2012: 11
FWCI
6.90
Type
journal article
Language
en
Open access
Locations
  • Chemistry & Biology (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • National Institutes of Health (5r21ns051214-02)
  • Canadian Institutes of Health Research (82944)
  • National Institute of Neurological Disorders and Stroke (5R21NS051214-02, R21 NS051214)
Topics
  • Health Sciences › Medicine › Physiology › Lysosomal Storage Disorders Research
  • Health Sciences › Medicine › Rheumatology › Glycogen Storage Diseases and Myoclonus
  • Physical Sciences › Chemistry › Organic Chemistry › Carbohydrate Chemistry and Synthesis
Keywords
pharmacological chaperones, beta-hexosaminidase A, Tay-Sachs disease, Sandhoff disease, competitive inhibitors, high-throughput screening, endoplasmic reticulum retention, lysosomal enzyme transport, patient fibroblasts
MeSH terms
Adult, beta-N-Acetylhexosaminidases, Dose-Response Relationship, Drug, Enzyme Inhibitors, Female, Fibroblasts, Humans, Indans, Lysosomes, Pyridazines, Sandhoff Disease, Tay-Sachs Disease, Molecular Chaperones, Inhibitory Concentration 50, Naphthalimides

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