{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,20]],"date-time":"2026-04-20T09:00:36Z","timestamp":1776675636112,"version":"3.51.2"},"reference-count":39,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2026,4,18]],"date-time":"2026-04-18T00:00:00Z","timestamp":1776470400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>GM1 gangliosidosis and Morquio B are rare lysosomal storage disorders (LSDs) with significant unmet medical needs. These disorders result from mutations in the galactosidase beta 1 (GLB1) gene, leading to impaired \u03b2-galactosidase (\u03b2-Gal) activity and toxic substrate accumulation. The lack of approved disease-modifying therapies for GM1 gangliosidosis and Morquio B, along with the challenges of achieving effective central nervous system delivery, has driven interest in small-molecule pharmacological chaperones (PCs) to restore \u03b2-Gal stability and function. Using Gain Therapeutics\u2019 Magellan\u2122 platform, a novel allosteric binding site on \u03b2-Gal was identified, enabling the discovery of a new class of Structurally Targeted Allosteric Regulators (STARs). Medicinal chemistry optimization produced a structurally unique STAR compound series, demonstrating broad \u03b2-Gal stabilizing effects. The therapeutic potential of these compounds was evaluated in vitro using a canine fibroblast model of GM1 gangliosidosis, where they were shown to significantly reduce toxic GM1 ganglioside accumulation. Immunocytochemistry-based assays confirmed substrate clearance and provided reliable structure\u2013activity relationships, guiding further compound development. Notably, STARs achieved greater substrate clearance than the competitive PC N-nonyl-deoxygalactonojirimycin (NN-DGJ) under the conditions tested, as demonstrated by immunocytochemistry-based assays. While these findings are encouraging, further in vivo studies are required to validate the therapeutic efficacy of these few STAR compounds, particularly in addressing the neurodegenerative aspects of GM1 gangliosidosis. This study underscores the potential of the Magellan platform in identifying STAR molecules and provides a strong foundation for further optimization and preclinical validation in GLB1-related disorders, particularly GM1 gangliosidosis.<\/jats:p>","DOI":"10.3390\/ijms27083631","type":"journal-article","created":{"date-parts":[[2026,4,20]],"date-time":"2026-04-20T08:24:23Z","timestamp":1776673463000},"page":"3631","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development of Small-Molecule Allosteric Modulators of Beta-Galactosidase (\u03b2-Gal) for the Treatment of GM1 Gangliosidosis and Morquio B"],"prefix":"10.3390","volume":"27","author":[{"given":"Nat\u00e0lia","family":"P\u00e9rez-Carmona","sequence":"first","affiliation":[{"name":"Gain Therapeutics Sucursal en Espa\u00f1a, Parc Cient\u00edfic de Barcelona, 08028 Barcelona, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2239-6670","authenticated-orcid":false,"given":"Elena","family":"Cubero","sequence":"additional","affiliation":[{"name":"Gain Therapeutics Sucursal en Espa\u00f1a, Parc Cient\u00edfic de Barcelona, 08028 Barcelona, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ana","family":"Ruano","sequence":"additional","affiliation":[{"name":"Gain Therapeutics Sucursal en Espa\u00f1a, Parc Cient\u00edfic de Barcelona, 08028 Barcelona, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5008-5741","authenticated-orcid":false,"given":"Maria","family":"Pons-Vizcarra","sequence":"additional","affiliation":[{"name":"Gain Therapeutics Sucursal en Espa\u00f1a, Parc Cient\u00edfic de Barcelona, 08028 Barcelona, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2793-1545","authenticated-orcid":false,"given":"Aida","family":"Delgado","sequence":"additional","affiliation":[{"name":"Gain Therapeutics Sucursal en Espa\u00f1a, Parc Cient\u00edfic de Barcelona, 08028 Barcelona, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4526-7895","authenticated-orcid":false,"given":"Ana","family":"Trapero","sequence":"additional","affiliation":[{"name":"Gain Therapeutics Sucursal en Espa\u00f1a, Parc Cient\u00edfic de Barcelona, 08028 Barcelona, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marc","family":"Reves","sequence":"additional","affiliation":[{"name":"Minoryx Therapeutics, 08302 Barcelona, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5631-1172","authenticated-orcid":false,"given":"Laura","family":"Rodr\u00edguez-Pascau","sequence":"additional","affiliation":[{"name":"Minoryx Therapeutics, 08302 Barcelona, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joanne","family":"Taylor","sequence":"additional","affiliation":[{"name":"Gain Therapeutics, Inc., Bethesda, MD 20814, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marc","family":"Martinell","sequence":"additional","affiliation":[{"name":"Minoryx Therapeutics, 08302 Barcelona, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0281-1347","authenticated-orcid":false,"given":"Xavier","family":"Barril","sequence":"additional","affiliation":[{"name":"Gain Therapeutics Sucursal en Espa\u00f1a, Parc Cient\u00edfic de Barcelona, 08028 Barcelona, Spain"},{"name":"Facultat de Farm\u00e0cia, IBUB & IQTC, Universitat de Barcelona, 08028 Barcelona, Spain"},{"name":"Catalan Institution for Research and Advanced Studies (ICREA), 08010 Barcelona, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-9582-7429","authenticated-orcid":false,"given":"Ana Mar\u00eda","family":"Garc\u00eda-Collazo","sequence":"additional","affiliation":[{"name":"Gain Therapeutics Sucursal en Espa\u00f1a, Parc Cient\u00edfic de Barcelona, 08028 Barcelona, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Nicoli, E.R., Annunziata, I., d\u2019Azzo, A., Platt, F.M., Tifft, C.J., and Stepien, K.M. (2021). GM1 gangliosidosis\u2014A mini-review. Front. Genet., 12.","DOI":"10.3389\/fgene.2021.734878"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1016\/S0925-4439(99)00075-7","article-title":"Molecular basis of GM1 gangliosidosis and Morquio disease, type B. Structure\u2013function studies of lysosomal \u03b2-galactosidase and the non-lysosomal \u03b2-galactosidase-like protein","volume":"1455","author":"Callahan","year":"1999","journal-title":"Biochim. Biophys. Acta Mol. Basis Dis."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"391","DOI":"10.1016\/j.ymgme.2008.04.012","article-title":"GM1 gangliosidosis: Review of clinical, molecular, and therapeutic aspects","volume":"94","author":"Scaglia","year":"2008","journal-title":"Mol. Genet. Metab."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1011","DOI":"10.1007\/s10545-005-0112-z","article-title":"Cumulative incidence rates of the mucopolysaccharidoses in Germany","volume":"28","author":"Baehner","year":"2005","journal-title":"J. Inherit. Metab. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Yuskiv, N., Higaki, K., and Stockler-Ipsiroglu, S. (2020). Morquio B disease. Disease characteristics and treatment options of a distinct GLB1-related dysostosis multiplex. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21239121"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Foster, D., Williams, L., Arnold, N., and Larsen, J. (2024). Therapeutic developments for neurodegenerative GM1 gangliosidosis. Front. Neurosci., 18.","DOI":"10.3389\/fnins.2024.1392683"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2980","DOI":"10.1002\/tcr.202100269","article-title":"Pharmacological chaperones for \u03b2-galactosidase related to G(M1)\u2014Gangliosidosis and Morquio B: Recent sdvances","volume":"21","author":"Thonhofer","year":"2021","journal-title":"Chem. Rec."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"262","DOI":"10.1016\/j.ymgme.2010.03.019","article-title":"DLHex-DGJ, a novel derivative of 1-deoxygalactonojirimycin with pharmacological chaperone activity in human G(M1)-gangliosidosis fibroblasts","volume":"100","author":"Fantur","year":"2010","journal-title":"Mol. Genet. Metab."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"10216","DOI":"10.1016\/j.bmc.2008.10.054","article-title":"1-Deoxygalactonojirimycin-lysine hybrids as potent D-galactosidase inhibitors","volume":"16","author":"Steiner","year":"2008","journal-title":"Bioorganic Med. Chem."},{"key":"ref_10","first-page":"7","article-title":"Chaperone therapy for neuronopathic lysosomal diseases: Competitive inhibitors as chemical chaperones for enhancement of mutant enzyme activities","volume":"3","author":"Suzuki","year":"2009","journal-title":"Perspect. Med. Chem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1089\/adt.2011.0370","article-title":"Identification and characterization of pharmacological chaperones to correct enzyme deficiencies in lysosomal storage disorders","volume":"9","author":"Valenzano","year":"2011","journal-title":"Assay Drug Dev. Technol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1002\/emmm.200900036","article-title":"Treating lysosomal storage diseases with pharmacological chaperones: From concept to clinics","volume":"1","author":"Parenti","year":"2009","journal-title":"EMBO Mol. Med."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Tran, M.L., G\u00e9nisson, Y., Ballereau, S., and Dehoux, C. (2020). Second-generation pharmacological chaperones: Beyond inhibitors. Molecules, 25.","DOI":"10.3390\/molecules25143145"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"a004507","DOI":"10.1101\/cshperspect.a004507","article-title":"Chemical and biological approaches for adapting proteostasis to ameliorate protein misfolding and aggregation diseases: Progress and prognosis","volume":"3","author":"Lindquist","year":"2011","journal-title":"Cold Spring Harb. Perspect. Biol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"15912","DOI":"10.1073\/pnas.2536657100","article-title":"Chemical chaperone therapy for brain pathology in GM1-gangliosidosis","volume":"100","author":"Matsuda","year":"2003","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1016\/j.ymgme.2012.06.007","article-title":"Evaluation of N-nonyl-deoxygalactonojirimycin as a pharmacological chaperone for human GM1 gangliosidosis leads to identification of a feline model suitable for testing enzyme enhancement therapy","volume":"107","author":"Rigat","year":"2012","journal-title":"Mol. Genet. Metab."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1016\/j.ejmech.2016.09.095","article-title":"(5aR)-5a-C-Pentyl-4-epi-isofagomine: A powerful inhibitor of lysosomal \u03b2-galactosidase and a remarkable chaperone for mutations associated with GM1-gangliosidosis and Morquio disease type B","volume":"126","author":"Front","year":"2017","journal-title":"Eur. J. Med. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Clemente, F., Mart\u00ednez-Bail\u00e9n, M., Matassini, C., Morrone, A., Falliano, S., Caciotti, A., Paoli, P., Goti, A., and Cardona, F. (2022). Synthesis of a new \u03b2-galactosidase inhibitor displaying pharmacological chaperone properties for GM1 gangliosidosis. Molecules, 27.","DOI":"10.3390\/molecules27134008"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"4398","DOI":"10.1021\/jo500328u","article-title":"Synthesis of 1,5-dideoxy-1,5-iminoribitol C-glycosides through a nitrone\u2013olefin cycloaddition domino strategy: Identification of pharmacological chaperones of mutant human lysosomal \u03b2-galactosidase","volume":"79","author":"Siriwardena","year":"2014","journal-title":"J. Org. Chem."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Fregno, I., P\u00e9rez-Carmona, N., Rudinskiy, M., Sold\u00e0, T., Bergmann, T.J., Ruano, A., Delgado, A., Cubero, E., Bellotto, M., and Garc\u00eda-Collazo, A.M. (2025). Allosteric modulation of GCase enhances lysosomal activity and reduces ER stress in GCase-related disorders. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26094392"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Montpeyo, M., P\u00e9rez-Carmona, N., Cubero, E., Delgado, A., Ruano, A., Carrillo, J., Bellotto, M., Martinez-Vicente, M., and Garcia-Collazo, A.M. (2024). Developing allosteric chaperones for GBA1-associated disorders-An integrated computational and experimental approach. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26010009"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1801","DOI":"10.1074\/jbc.M111.293795","article-title":"Crystal structure of human \u03b2-galactosidase: Structural basis of GM1 gangliosidosis and Morquio B diseases","volume":"287","author":"Ohto","year":"2012","journal-title":"J. Biol. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Rudinskiy, M., Pons-Vizcarra, M., Sold\u00e0, T., Fregno, I., Bergmann, T.J., Ruano, A., Delgado, A., Morales, S., Barril, X., and Bellotto, M. (2023). Validation of a highly sensitive HaloTag-based assay to evaluate the potency of a novel class of allosteric \u03b2-galactosidase correctors. PLoS ONE, 18.","DOI":"10.1371\/journal.pone.0294437"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2363","DOI":"10.1021\/jm801385d","article-title":"Binding site detection and druggability index from first principles","volume":"52","author":"Seco","year":"2009","journal-title":"J. Med. Chem."},{"key":"ref_25","unstructured":"Barril Alonso, X., Alvarez Garcia, D., and Schmidtke, P. (2025, October 06). Method of Binding Site and Binding Energy Determination by Mixed Explicit Solvent Simulations. Available online: https:\/\/patentscope.wipo.int\/search\/en\/WO2013092922."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"8530","DOI":"10.1021\/jm5010418","article-title":"Molecular simulations with solvent competition quantify water displaceability and provide accurate interaction maps of protein binding sites","volume":"57","author":"Barril","year":"2014","journal-title":"J. Med. Chem."},{"key":"ref_27","first-page":"62","article-title":"Extracting atomic contributions to binding free energy using Molecular Dynamics Simulations with Mixed Solvents (MDmix)","volume":"19","author":"Schmidtke","year":"2022","journal-title":"Curr. Drug Discov. Technol."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Ruiz-Carmona, S., Alvarez-Garcia, D., Foloppe, N., Garmendia-Doval, A.B., Juhos, S., Schmidtke, P., Barril, X., Hubbard, R.E., and Morley, S.D. (2014). rDock: A fast, versatile and open source program for docking ligands to proteins and nucleic acids. PLoS Comput. Biol., 10.","DOI":"10.1371\/journal.pcbi.1003571"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1273","DOI":"10.1021\/ci010132r","article-title":"Reoptimization of MDL keys for use in drug discovery","volume":"42","author":"Durant","year":"2002","journal-title":"J. Chem. Inf. Comput. Sci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2212","DOI":"10.1038\/nprot.2007.321","article-title":"The use of differential scanning fluorimetry to detect ligand interactions that promote protein stability","volume":"2","author":"Niesen","year":"2007","journal-title":"Nat. Protoc."},{"key":"ref_31","unstructured":"Gomes, C.M. (2019). Thermal shift and stability assays of disease-related misfolded proteins using differential scanning fluorimetry. Protein Misfolding Diseases: Methods and Protocols, Springer."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"593","DOI":"10.1023\/A:1005630013448","article-title":"Isolation and characterization of the normal canine beta-galactosidase gene and its mutation in a dog model of GM1-gangliosidosis","volume":"23","author":"Wang","year":"2000","journal-title":"J. Inherit. Metab. Dis."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"541","DOI":"10.1146\/annurev.biophys.26.1.541","article-title":"Use of surface plasmon resonance to probe the equilibrium and dynamic aspects of interactions between biological macromolecules","volume":"26","author":"Schuck","year":"1997","journal-title":"Annu. Rev. Biophys. Biomol. Struct."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1417","DOI":"10.1016\/j.neuroscience.2006.01.064","article-title":"Molecular identification of an N-type Ca2+ channel in saccular hair cells","volume":"139","author":"Ramakrishnan","year":"2006","journal-title":"Neuroscience"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"3431","DOI":"10.1016\/j.bmcl.2017.05.086","article-title":"A new type of pharmacological chaperone for G(M1)-gangliosidosis related human lysosomal \u03b2-galactosidase: N-Substituted 5-amino-1-hydroxymethyl-cyclopentanetriols","volume":"27","author":"Schalli","year":"2017","journal-title":"Bioorganic Med. Chem. Lett."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Edelmann, M.J., and Maegawa, G.H.B. (2020). CNS-targeting therapies for lysosomal storage diseases: Current advances and challenges. Front. Mol. Biosci., 7.","DOI":"10.3389\/fmolb.2020.559804"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"874","DOI":"10.1002\/jimd.12615","article-title":"Advances in therapies for neurological lysosomal storage disorders","volume":"46","author":"Ellison","year":"2023","journal-title":"J. Inh. Metab. Dis."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"366","DOI":"10.1002\/minf.201000019","article-title":"Bioisosteric replacement and scaffold hopping in lead generation and optimization","volume":"29","author":"Langdon","year":"2010","journal-title":"Mol. Inf."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1016\/j.drudis.2006.12.003","article-title":"Scaffold selection and scaffold hopping in lead generation: A medicinal chemistry perspective","volume":"12","author":"Zhao","year":"2007","journal-title":"Drug Discov. Today"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/8\/3631\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,20]],"date-time":"2026-04-20T08:33:31Z","timestamp":1776674011000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/8\/3631"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,18]]},"references-count":39,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2026,4]]}},"alternative-id":["ijms27083631"],"URL":"https:\/\/doi.org\/10.3390\/ijms27083631","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4,18]]}}}