High-Throughput Screening for Human Lysosomal β-N-Acetyl Hexosaminidase Inhibitors Acting as Pharmacological Chaperones
Open accessAbstract
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.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
The Metabolic and Molecular Bases of Inherited Disease
Scriver, R. C. (1995) · Medical Entomology and Zoology · Cited by 12,101 · see citing works
Zhang, J.-H., Chung, T. D. Y., & Oldenburg, K. R. (1999) · SLAS Discovery · doi:10.1177/108705719900400206 · Cited by 6,847 · see citing works
Structural and sequence-based classification of glycoside hydrolases
Henrissat, B., & Davies, G. J. (1997) · Current Opinion in Structural Biology · doi:10.1016/s0959-440x(97)80072-3 · Cited by 1,668 · see citing works
Hasilík, A., & Neufeld, E. F. (1980) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)85585-0 · Cited by 803 · see citing works
Fan, J., Ishii, S., Asano, N., & Suzuki, Y. (1999) · Nature Medicine · doi:10.1038/4801 · Cited by 616 · see citing works
Sawkar, A. R., Cheng, W., Beutler, E., Wong, C., Balch, W. E., & Kelly, J. W. (2002) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.192582899 · Cited by 508 · see citing works
Rye, C. S., & Withers, S. G. (2000) · Current Opinion in Chemical Biology · doi:10.1016/s1367-5931(00)00135-6 · Cited by 470 · see citing works
Self-citationBiosynthesis of lysosomal enzymes in fibroblasts. Phosphorylation of mannose residues
Hasilik, A., & Neufeld, E. F. (1980) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)85586-2 · Cited by 400 · see citing works
O-GlcNAcase Uses Substrate-assisted Catalysis
Macauley, M. S., Whitworth, G. E., Debowski, A. W., Chin, D., & Vocadlo, D. J. (2005) · Journal of Biological Chemistry · doi:10.1074/jbc.m413819200 · Cited by 378 · see citing works
Du, K., Sharma, M., & Lukacs, G. L. (2005) · Nature Structural & Molecular Biology · doi:10.1038/nsmb882 · Cited by 356 · see citing works
Tews, I., Anastassis Perrakis, A., Oppenheim, A. B., Dauter, Z., Wilson, K. S., & Vorgias, C. E. (1996) · Nature Structural Biology · doi:10.1038/nsb0796-638 · Cited by 342 · see citing works
Pharmacological chaperones: potential treatment for conformational diseases
Bernier, V., Lagacé, M., Bichet, D. G., & Bouvier, M. (2004) · Trends in Endocrinology and Metabolism · doi:10.1016/j.tem.2004.05.003 · Cited by 285 · see citing works
Naphthalimides as Anticancer Agents: Synthesis and Biological Activity
BRANA, M. F., & Ramos, A. (2001) · Anti-Cancer Agents in Medicinal Chemistry · doi:10.2174/1568011013354624 · Cited by 282 · see citing works
Friedler, A., Hansson, L. O., Veprintsev, D. B., Freund, S. M. V., Rippin, T. M., Nikolova, P. V., Proctor, M. R., Rüdiger, S., & Fersht, A. R. (2002) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.241629998 · Cited by 268 · see citing works
Chemical chaperone therapy for brain pathology in G M1 -gangliosidosis
Matsuda, J., Suzuki, O., Oshima, A., et al. (2003) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.2536657100 · Cited by 264 · see citing works
Crystallographic Evidence for Substrate-assisted Catalysis in a Bacterial β-Hexosaminidase
Mark, B. L., Vocadlo, D. J., Knapp, S., Triggs‐Raine, B. L., Withers, S. G., & James, M. N. G. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m011067200 · Cited by 257 · see citing works
Self-citationUlloa‐Aguirre, A., Janovick, J. A., Brothers, S. P., & Conn, P. M. (2004) · Traffic · doi:10.1111/j.1600-0854.2004.00232.x · Cited by 252 · see citing works
NAG-thiazoline, An N-Acetyl-β-hexosaminidase Inhibitor That Implicates Acetamido Participation
Knapp, S., Vocadlo, D. J., Gao, Z., Kirk, B. A., Lou, J., & Withers, S. G. (1996) · Journal of the American Chemical Society · doi:10.1021/ja960826u · Cited by 248 · see citing works
Self-citationAsano, N., Ishii, S., Kizu, H., et al. (2000) · European Journal of Biochemistry · doi:10.1046/j.1432-1327.2000.01457.x · Cited by 245 · see citing works
A contradictory treatment for lysosomal storage disorders: inhibitors enhance mutant enzyme activity
Fan, J. (2003) · Trends in Pharmacological Sciences · doi:10.1016/s0165-6147(03)00158-5 · Cited by 232 · see citing works
Mark, B. L., Mahuran, D. J., Cherney, M. M., Zhao, D., Knapp, S., & James, M. N. G. (2003) · Journal of Molecular Biology · doi:10.1016/s0022-2836(03)00216-x · Cited by 224 · see citing works
Self-citationEnzymes of Lipid Metabolism II
Freysz, L., Dreyfus, H., Massarelli, R., et al. (1986) · Springer eBooks · doi:10.1007/978-1-4684-5212-9 · Cited by 206 · see citing works
High-Throughput Screening Identifies Inhibitors of the SARS Coronavirus Main Proteinase
Blanchard, J., Elowe, N. H., Huitema, C., Fortin, P. D., Cechetto, J. D., Eltis, L. D., & Brown, E. D. (2004) · Chemistry & Biology · doi:10.1016/j.chembiol.2004.08.011 · Cited by 205 · see citing works
Self-citationSelective Optimization of Side Activities: Another Way for Drug Discovery
Wermuth, C. G. (2004) · Journal of Medicinal Chemistry · doi:10.1021/jm030480f · Cited by 202 · see citing works
Tropak, M. B., Reid, S. P., Guiral, M., et al. (2004) · Journal of Biological Chemistry · doi:10.1074/jbc.m308523200 · Cited by 200 · see citing works
Self-citationLemieux, M. J., Mark, B. L., Cherney, M. M., et al. (2006) · Journal of Molecular Biology · doi:10.1016/j.jmb.2006.04.004 · Cited by 195 · see citing works
Gaucher Disease-Associated Glucocerebrosidases Show Mutation-Dependent Chemical Chaperoning Profiles
Sawkar, A. R., Adamski‐Werner, S. L., Cheng, W., Wong, C., Beutler, E., Zimmer, K., & Kelly, J. W. (2005) · Chemistry & Biology · doi:10.1016/j.chembiol.2005.09.007 · Cited by 182 · see citing works
Ciechanover, A. J. (2005) · Angewandte Chemie International Edition · doi:10.1002/anie.200501428 · Cited by 180 · see citing works
Yam, G. H., Zuber, C., & Roth, J. (2005) · The FASEB Journal · doi:10.1096/fj.04-2375com · Cited by 163 · see citing works
Charria-Ortiz, G. A. (2007) · Springer eBooks · doi:10.1007/978-0-387-70909-3_16 · Cited by 132 · see citing works
Pharmacological chaperone action on G-protein-coupled receptors
Bernier, V., Bichet, D. G., & Bouvier, M. (2004) · Current Opinion in Pharmacology · doi:10.1016/j.coph.2004.08.001 · Cited by 130 · see citing works
The X-ray Crystal Structure of Human β-Hexosaminidase B Provides New Insights into Sandhoff Disease
Maier, T., Sträter, N., Schuette, C. G., Klingenstein, R., Sandhoff, K., & Saenger, W. (2003) · Journal of Molecular Biology · doi:10.1016/s0022-2836(03)00311-5 · Cited by 116 · see citing works
Liang, P., Cheng, W., Lee, Y.-L., Yu, H., Wu, Y., Lin, Y., & Wong, C. (2006) · ChemBioChem · doi:10.1002/cbic.200500321 · Cited by 109 · see citing works
Rescue of Vasopressin V2 Receptor Mutants by Chemical Chaperones: Specificity and Mechanism
Robben, J. H., Sze, M., Knoers, N. V. A. M., & Deen, P. M. T. (2005) · Molecular Biology of the Cell · doi:10.1091/mbc.e05-06-0579 · Cited by 99 · see citing works
The biochemistry of HEXA and HEXB gene mutations causing GM2 gangliosidosis
Mahuran, D. J. (1991) · Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease · doi:10.1016/0925-4439(91)90044-a · Cited by 98 · see citing works
Self-citationApplication of High-Throughput, Molecular-Targeted Screening to Anticancer Drug Discovery
Shoemaker, R., Scudiero, D. A., Melillo, G., Currens, M. J., Monks, A. P., Rabow, A. A., Covell, D. G., & Sausville, E. A. (2002) · Current Topics in Medicinal Chemistry · doi:10.2174/1568026023394317 · Cited by 91 · see citing works
Diettrich, O., Mills, K. A., Johnson, A., Hasilík, A., & WINCHESTER, B. G. (1998) · FEBS Letters · doi:10.1016/s0014-5793(98)01578-6 · Cited by 83 · see citing works
Liu, J., Numa, M. M. D., Liu, H., et al. (2004) · The Journal of Organic Chemistry · doi:10.1021/jo049355h · Cited by 77 · see citing works
Hou, Y., Tse, R., & Mahuran, D. J. (1996) · Biochemistry · doi:10.1021/bi9524575 · Cited by 70 · see citing works
Self-citationZolli‐Juran, M., Cechetto, J. D., Hartlen, R., et al. (2003) · Bioorganic & Medicinal Chemistry Letters · doi:10.1016/s0960-894x(03)00480-3 · Cited by 69 · see citing works
Tews, I., Vincentelli, R., & Vorgias, C. E. (1996) · Gene · doi:10.1016/0378-1119(95)00848-9 · Cited by 68 · see citing works
Faulty association of alpha- and beta-subunits in some forms of beta-hexosaminidase A deficiency.
d’Azzo, A., Proia, R. L., Kolodny, E. H., et al. (1984) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)90623-x · Cited by 66 · see citing works
Bis-naphthalimides: a new class of antitumor agents.
BRANA, M. F., Castellano, J. M., Morán, M., de Vega, M. J. P., Romerdahl, C., Qian, X.-D., Bousquet, P. F., Emling, F., Schlick, E., & Keilhauer, G. (1993) · PubMed · Cited by 55 · see citing works
Biochemical Basis of Late-Onset Neurolipidoses
Conzelmann, E., & Sandhoff, K. (1991) · Developmental Neuroscience · doi:10.1159/000112160 · Cited by 52 · see citing works
Mechanisms of Dimethyl Sulfoxide Augmentation of IL-1β Production
Xing, L., & Remick, D. G. (2005) · The Journal of Immunology · doi:10.4049/jimmunol.174.10.6195 · Cited by 48 · see citing works
Kappes, E., & Legler, G. (1989) · Journal of Carbohydrate Chemistry · doi:10.1080/07328308908048567 · Cited by 48 · see citing works
Brown, C. A., & Mahuran, D. J. (1993) · PubMed · Cited by 43 · see citing works
Self-citationAldose Reductase Inhibitors: An Update
Tsai, S. C., & Burnakis, T. G. (1993) · Annals of Pharmacotherapy · doi:10.1177/106002809302700616 · Cited by 40 · see citing works
Kakkar, T., Pak, Y., & Mayersohn, M. (2000) · Journal of Pharmacology and Experimental Therapeutics · doi:10.1016/s0022-3565(24)39308-5 · Cited by 37 · see citing works
Villalona-Calero, M. A., Eder, J. P., Toppmeyer, D., et al. (2001) · Journal of Clinical Oncology · doi:10.1200/jco.2001.19.3.857 · Cited by 37 · see citing works
Tay-Sachs disease with hexosaminidase A: characterization of the defective enzyme in two patients.
Bayleran, J. K., Hechtman, P., Kolodny, E. H., et al. (1987) · PubMed · Cited by 26 · see citing works
Translation initiation in the HEXB gene encoding the beta-subunit of human beta-hexosaminidase.
Neote, K. S., Brown, C. A., Mahuran, D. J., et al. (1990) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(17)45286-0 · Cited by 26 · see citing works
Self-citationHou, Y., McInnes, B., Hinek, A., et al. (1998) · Journal of Biological Chemistry · doi:10.1074/jbc.273.33.21386 · Cited by 26 · see citing works
GM1-gangliosidosis (Type II): Studies on a fibroblast cell strain
Callahan, J. W., Pinsky, L., & Wolfe, L. S. (1970) · Biochemical Medicine · doi:10.1016/0006-2944(70)90056-6 · Cited by 23 · see citing works
A Decade of Developments in Diuretic Drug Therapy
Hutcheon, D. E., & Martinez, J. C. (1986) · The Journal of Clinical Pharmacology · doi:10.1002/j.1552-4604.1986.tb02951.x · Cited by 17 · see citing works
Awada, A. H., Thödtmann, R., Piccart, M. J., et al. (2003) · European Journal of Cancer · doi:10.1016/s0959-8049(02)00776-1 · Cited by 15 · see citing works
A continuous fluorimetric assay for glycosidase activity: Human N-acetyl-β-d-hexosaminidase
Shulman, M. L., Kulshin, V. A., & Khorlin, A. Y. (1980) · Analytical Biochemistry · doi:10.1016/0003-2697(80)90198-0 · Cited by 9 · see citing works
Molecular Heterogeneity in O-Variant GM2 Gangliosidosis
O’Dowd, B. F., Klavins, M. H., Willard, H. F., et al. (1986) · Springer eBooks · doi:10.1007/978-1-4684-5212-9_92 · Cited by 2 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
De Masi, R., & Orlando, S. (2022) · International Journal of Molecular Sciences · doi:10.3390/ijms23137373 · Cited by 20 · see citing works
“A promising therapy for Tay–Sachs and Sandhoff diseases involves the use of β-Nacetylhexosaminidase inhibitors such as 2-acetamido-1,4-imino-1,2,4-tride-oxy-l-arabinitol (LABNAc) as a chemical chaperone to enhance the enzyme activity above subcritical levels in order to avoid glycolipid storage [237].”
Background
- 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)
- PMC
- PMC1989145
- 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
- bronze
- doi.org
- Licence
- 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
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review