{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T14:51:28Z","timestamp":1790520688711,"version":"4.1.0"},"reference-count":55,"publisher":"American Chemical Society (ACS)","issue":"3","license":[{"start":{"date-parts":[[2019,1,24]],"date-time":"2019-01-24T00:00:00Z","timestamp":1548288000000},"content-version":"vor","delay-in-days":14,"URL":"https:\/\/pubs.acs.org\/page\/policy\/authorchoice_ccbyncnd_termsofuse.html"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2019,3,20]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Cholera is a potentially fatal bacterial infection that affects a large number of people in developing countries. It is caused by the cholera toxin (CT), an AB5 toxin secreted by Vibrio cholera. The toxin comprises a toxic A-subunit and a pentameric B-subunit that bind to the intestinal cell surface. Several monovalent and multivalent inhibitors of the toxin have been synthesized but are too complicated and expensive for practical use in developing countries. Meta-nitrophenyl ?-galactoside (MNPG) is a known promising ligand for CT, and here mono- and multivalent compounds based on MNPG were synthesized. We present the synthesis of MNPG in greatly improved yields and its use while linked to a multivalent scaffold. We used economical polymers as multivalent scaffolds, namely, polyacrylamide, dextran, and hyperbranched polyglycerols (hPGs). Copper-catalyzed alkyne azide cycloaddition reaction (CuAAC) produced the inhibitors that were tested in an ELISA-type assay and an intestinal organoid swelling inhibition assay. The inhibitory properties varied widely depending on the type of polymer, and the most potent conjugates showed IC50 values in the nanomolar range.<\/jats:p>","DOI":"10.1021\/acs.bioconjchem.8b00902","type":"journal-article","created":{"date-parts":[[2019,1,10]],"date-time":"2019-01-10T12:24:41Z","timestamp":1547123081000},"page":"785-792","source":"Crossref","is-referenced-by-count":28,"title":["Strong Inhibition of Cholera Toxin B Subunit by Affordable,\nPolymer-Based Multivalent Inhibitors"],"prefix":"10.1021","volume":"30","author":[{"given":"Diksha","family":"Haksar","sequence":"first","affiliation":[{"name":"Utrecht University , , Universiteitsweg 99 , ,","place":["Utrecht, The Netherlands, 3584 CG"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eyleen","family":"de Poel","sequence":"additional","affiliation":[{"name":"University Medical Centre Utrecht , , Lundlaan 6 , ,","place":["Utrecht, The Netherlands, 3508 GA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Linda Quarles","family":"van Ufford","sequence":"additional","affiliation":[{"name":"Utrecht University , , Universiteitsweg 99 , ,","place":["Utrecht, The Netherlands, 3584 CG"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sumati","family":"Bhatia","sequence":"additional","affiliation":[{"name":"Freie Universita?t at Berlin , , Takustr. 3 , ,","place":["Berlin, Germany, 14195"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rainer","family":"Haag","sequence":"additional","affiliation":[{"name":"Freie Universita?t at Berlin , , Takustr. 3 , ,","place":["Berlin, Germany, 14195"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeffrey","family":"Beekman","sequence":"additional","affiliation":[{"name":"University Medical Centre Utrecht , , Lundlaan 6 , ,","place":["Utrecht, The Netherlands, 3508 GA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4723-3584","authenticated-orcid":true,"given":"Roland J.","family":"Pieters","sequence":"additional","affiliation":[{"name":"Utrecht University , , Universiteitsweg 99 , ,","place":["Utrecht, The Netherlands, 3584 CG"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2019,1,10]]},"reference":[{"key":"2026082516542544900_cit1","unstructured":"World Health Organization.\n          \n          https:\/\/www.who.int\/en\/news-room\/fact-sheets\/detail\/cholera (accessed Jan. 9, 2019)."},{"key":"2026082516542544900_cit2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0003832","article-title":"Updated global burden\nof cholera in endemic countries","volume":"9","author":"Ali","year":"2015","journal-title":"PLoS Neglected\nTrop. Dis."},{"key":"#cr-split#-2026082516542544900_cit3.1","unstructured":"WHO Annual Report 2017 Yemen"},{"key":"#cr-split#-2026082516542544900_cit3.2","unstructured":"WHO Regional Office for the Eastern Mediterranean: Cairo, 2018. Cairo. License: CC BYNC-SA 3.0 IGO."},{"key":"2026082516542544900_cit4","doi-asserted-by":"publisher","first-page":"10","DOI":"10.1186\/s12941-017-0185-1","article-title":"Pandemics, pathogenicity\nand changing molecular epidemiology of cholera in the era of global\nwarming","volume":"16","author":"Chowdhury","year":"2017","journal-title":"Ann. Clin. Microbiol. Antimicrob."},{"key":"2026082516542544900_cit5","first-page":"638","article-title":"Vaxchora: The First\nFDA-Approved Cholera Vaccination in the United\nStates","volume":"42","author":"Mosley","year":"2017","journal-title":"P T"},{"key":"2026082516542544900_cit6","first-page":"78","article-title":"Monovalent and Multivalent Inhibitors of Bacterial Toxins","volume-title":"Synthesis and Biological Applications of Glycoconjugates","author":"Turnbull","year":"2011"},{"key":"2026082516542544900_cit7","doi-asserted-by":"publisher","first-page":"974","DOI":"10.3390\/toxins7030974","article-title":"Cholera toxin B: one\nsubunit with many pharmaceutical applications","volume":"7","author":"Baldauf","year":"2015","journal-title":"Toxins"},{"key":"2026082516542544900_cit8","doi-asserted-by":"publisher","first-page":"274","DOI":"10.1021\/acs.accounts.5b00480","article-title":"Fighting Cholera\nOne-on-One: The Development and E ffi cacy of Multivalent Cholera-Toxin-Binding\nMolecules","volume":"49","author":"Zuilhof","year":"2016","journal-title":"Acc. Chem. Res."},{"key":"2026082516542544900_cit9","doi-asserted-by":"publisher","first-page":"484","DOI":"10.3762\/bjoc.14.34","article-title":"Carbohydrate inhibitors\nof cholera toxin","volume":"14","author":"Kumar","year":"2018","journal-title":"Beilstein J. Org. Chem."},{"key":"2026082516542544900_cit10","doi-asserted-by":"publisher","first-page":"1047","DOI":"10.1021\/ja0378207","article-title":"Dissecting\nthe Cholera Toxin-Ganglioside\nGM1 Interaction by Isothermal Titration Calorimetry","volume":"126","author":"Turnbull","year":"2004","journal-title":"J. Am. Chem. Soc."},{"key":"2026082516542544900_cit11","doi-asserted-by":"publisher","first-page":"758","DOI":"10.1021\/acsinfecdis.7b00085","article-title":"Fucosylated Molecules\nCompetitively Interfere with Cholera Toxin Binding to Host Cells","volume":"4","author":"Wands","year":"2018","journal-title":"ACS Infect. Dis."},{"key":"2026082516542544900_cit12","doi-asserted-by":"publisher","first-page":"1","DOI":"10.7554\/eLife.09545","article-title":"Fucosylation and\nprotein glycosylation create functional receptors for cholera toxin","volume":"4","author":"Wands","year":"2015","journal-title":"eLife"},{"key":"2026082516542544900_cit13","doi-asserted-by":"publisher","first-page":"2032","DOI":"10.1021\/ja983567c","article-title":"Sugar mimics:\nAn artificial receptor for cholera toxin","volume":"121","author":"Bernardi","year":"1999","journal-title":"J.\nAm. Chem. Soc."},{"key":"2026082516542544900_cit14","doi-asserted-by":"publisher","first-page":"2197","DOI":"10.1016\/S0960-894X(00)00428-5","article-title":"Second Generation\nMimics of Ganglioside GM1 as Arti?cial Receptors for Cholera\nToxin: Replacement of the Sialic Acid Moiety","volume":"10","author":"Bernardi","year":"2000","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"2026082516542544900_cit15","doi-asserted-by":"publisher","first-page":"3660","DOI":"10.1021\/ja0444029","article-title":"A Synthetic\nDivalent\nCholera Toxin Glycocalix[4]arene Ligand Having Higher Affinity than\nNatural GM1 Oligosaccharide Scheme 1. Synthesis of the Divalent Ligand\n1","volume":"127","author":"Arosio","year":"2005","journal-title":"J. Am. Chem. Soc."},{"key":"2026082516542544900_cit16","doi-asserted-by":"publisher","first-page":"1951","DOI":"10.1002\/chem.200902469","article-title":"Synthesis and affinity\nevaluation of a small library of bidentate cholera toxin ligands:\nTowards nonhydrolyzable ganglioside mimics","volume":"16","author":"Cheshev","year":"2010","journal-title":"Chem. - Eur. J."},{"key":"2026082516542544900_cit17","doi-asserted-by":"publisher","first-page":"14868","DOI":"10.1039\/C3RA47500J","article-title":"Synthesis of RAFT polymers as bivalent inhibitors of\ncholera toxin","volume":"4","author":"Leaver","year":"2014","journal-title":"RSC Adv."},{"key":"2026082516542544900_cit18","doi-asserted-by":"publisher","first-page":"8465","DOI":"10.1039\/c1ob06317k","article-title":"Synthesis\nof 1,2,3-triazole linked galactopyranosides and evaluation of cholera\ntoxin inhibition","volume":"9","author":"Leaver","year":"2011","journal-title":"Org. Biomol. Chem."},{"key":"2026082516542544900_cit19","doi-asserted-by":"publisher","first-page":"2113","DOI":"10.1039\/b405344c","article-title":"Synthesis and cholera\ntoxin binding properties of multivalent\nGM1 mimics","volume":"2","author":"Arosio","year":"2004","journal-title":"Org. Biomol. Chem."},{"key":"2026082516542544900_cit20","doi-asserted-by":"publisher","first-page":"7812","DOI":"10.1002\/anie.201202945","article-title":"Probing bacterial-toxin inhibition\nwith synthetic glycopolymers\nprepared by tandem post-polymerization modification: role of linker\nlength and carbohydrate density","volume":"51","author":"Richards","year":"2012","journal-title":"Angew. Chem.,\nInt. Ed."},{"key":"2026082516542544900_cit21","doi-asserted-by":"publisher","first-page":"483","DOI":"10.1021\/bm050672n","article-title":"Effects of polymer structure on the inhibition of cholera\ntoxin by linear polypeptide-based glycopolymers","volume":"7","author":"Polizzotti","year":"2006","journal-title":"Biomacromolecules"},{"key":"2026082516542544900_cit22","doi-asserted-by":"publisher","first-page":"47","DOI":"10.1016\/j.carres.2016.05.011","article-title":"Synthetic multivalent ligands for\ncholera & cholera-like toxins: Protected cyclic neoglycopeptides","volume":"431","author":"Kumar","year":"2016","journal-title":"Carbohydr. Res."},{"key":"2026082516542544900_cit23","doi-asserted-by":"publisher","first-page":"8323","DOI":"10.1002\/anie.201404397","article-title":"A protein-based\npentavalent inhibitor of the cholera toxin B-subunit","volume":"53","author":"Branson","year":"2014","journal-title":"Angew. Chem., Int. 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Chem."},{"key":"2026082516542544900_cit26","doi-asserted-by":"publisher","first-page":"1500","DOI":"10.1002\/cbic.200700266","article-title":"Strong Inhibition of Cholera Toxin by Multivalent GM1\nDerivatives","volume":"8","author":"Pukin","year":"2007","journal-title":"ChemBioChem"},{"key":"2026082516542544900_cit27","doi-asserted-by":"publisher","first-page":"e0006266","DOI":"10.1371\/journal.pntd.0006266","article-title":"Neutralization of cholera toxin with\nnanoparticle decoys for treatment of cholera","volume":"12","author":"Das","year":"2018","journal-title":"PLoS Neglected Trop. Dis."},{"key":"2026082516542544900_cit28","doi-asserted-by":"publisher","first-page":"81","DOI":"10.1016\/j.addr.2015.05.011","article-title":"Polymer antidotes\nfor toxin sequestration","volume":"90","author":"Weisman","year":"2015","journal-title":"Adv. Drug Delivery\nRev."},{"key":"2026082516542544900_cit29","doi-asserted-by":"publisher","first-page":"3658","DOI":"10.1039\/c0ob01089h","article-title":"Multifunctional\nmultivalency: a focused library of polymeric cholera toxin antagonists","volume":"9","author":"Tran","year":"2011","journal-title":"Org. Biomol. Chem."},{"key":"2026082516542544900_cit30","doi-asserted-by":"publisher","first-page":"1611","DOI":"10.1039\/C3SC52982G","article-title":"Glycopolymers with secondary binding\nmotifs mimic glycan branching and display bacterial lectin selectivity\nin addition to affinity","volume":"5","author":"Jones","year":"2014","journal-title":"Chem. Sci."},{"key":"2026082516542544900_cit31","doi-asserted-by":"crossref","first-page":"884","DOI":"10.1128\/jcm.21.6.884-890.1985","article-title":"In Vitro and In\nVivo Cholera Toxin Production by Classical\nand El Tor Isolates of Vibrio cholerae","volume":"21","author":"Turnbull","year":"1985","journal-title":"J. Clin.\nMicrobiol."},{"key":"2026082516542544900_cit32","doi-asserted-by":"publisher","first-page":"5684","DOI":"10.1021\/bi982649a","article-title":"Structure-based\nexploration of the ganglioside GM1 binding sites of Escherichia coli\nheat-labile enterotoxin and cholera toxin for the discovery of receptor\nantagonists","volume":"38","author":"Minke","year":"1999","journal-title":"Biochemistry"},{"key":"2026082516542544900_cit33","doi-asserted-by":"publisher","first-page":"215","DOI":"10.1016\/S1074-5521(02)00097-2","article-title":"Anchor-based design\nof improved cholera\ntoxin and E. coli heat-labile enterotoxin receptor binding antagonists\nthat display multiple binding modes","volume":"9","author":"Pickens","year":"2002","journal-title":"Chem. Biol."},{"key":"2026082516542544900_cit34","doi-asserted-by":"publisher","first-page":"907","DOI":"10.1016\/j.bmc.2003.12.019","article-title":"3,5-Substituted\nphenyl galactosides as leads in designing effective cholera toxin\nantagonists: Synthesis and crystallographic studies","volume":"12","author":"Mitchell","year":"2004","journal-title":"Bioorg. Med. Chem."},{"key":"2026082516542544900_cit35","doi-asserted-by":"publisher","first-page":"1377","DOI":"10.1021\/ol049835v","article-title":"Solution-\nand Solid-Phase\nSyntheses of Guanidine-Bridged, Water-Soluble Linkers for Multivalent\nLigand Design","volume":"6","author":"Zhang","year":"2004","journal-title":"Org. Lett."},{"key":"2026082516542544900_cit36","doi-asserted-by":"publisher","first-page":"5851","DOI":"10.1039\/b813414f","article-title":"Modular synthesis of\nmultivalent glycoarchitectures and their unique selectin binding behavior","author":"Papp","year":"2008","journal-title":"Chem. Commun."},{"key":"2026082516542544900_cit37","doi-asserted-by":"publisher","first-page":"190","DOI":"10.1002\/adma.200902144","article-title":"Dendritic polyglycerols\nfor biomedical applications","volume":"22","author":"Caldero?n","year":"2010","journal-title":"Adv. Mater."},{"key":"2026082516542544900_cit38","doi-asserted-by":"publisher","first-page":"2954","DOI":"10.1021\/ja994363e","article-title":"An approach\nto glycerol dendrimers\nand pseudo-dendritic polyglycerols [6]","volume":"122","author":"Haag","year":"2000","journal-title":"J. Am.\nChem. Soc."},{"key":"2026082516542544900_cit39","doi-asserted-by":"publisher","first-page":"22","DOI":"10.1016\/j.biomaterials.2017.05.028","article-title":"Linear polysialoside outperforms\ndendritic analogs for inhibition of influenza virus infection in vitro\nand in vivo","volume":"138","author":"Bhatia","year":"2017","journal-title":"Biomaterials"},{"key":"2026082516542544900_cit40","doi-asserted-by":"publisher","first-page":"7540","DOI":"10.1002\/anie.200901583","article-title":"Biocompatible\nFunctionalized Polyglycerol Microgels with Cell Penetrating\nProperties","volume":"48","author":"Sisson","year":"2009","journal-title":"Angew. Chem., Int. Ed."},{"key":"2026082516542544900_cit41","doi-asserted-by":"publisher","first-page":"6968","DOI":"10.1021\/acs.jmedchem.6b00770","article-title":"Functional\nCharacterization of Cholera Toxin Inhibitors Using Human Intestinal\nOrganoids","volume":"59","author":"Zomer-van Ommen","year":"2016","journal-title":"J. Med. Chem."},{"key":"2026082516542544900_cit42","doi-asserted-by":"publisher","first-page":"199","DOI":"10.1016\/0008-6215(86)80016-7","article-title":"Streptococcus\npneumoniae type XIV polysaccharide: synthesis\nof a repeating branched tetrasaccharide with dioxa-type spacer-arms","volume":"150","author":"Amvam-Zollo","year":"1986","journal-title":"Carbohydr. Res."},{"key":"2026082516542544900_cit43","doi-asserted-by":"publisher","first-page":"1449","DOI":"10.1002\/adma.201202881","article-title":"Modular\n?click-in-emulsion? bone-targeted nanogels","volume":"25","author":"Heller","year":"2013","journal-title":"Adv. Mater."},{"key":"2026082516542544900_cit44","doi-asserted-by":"publisher","first-page":"78","DOI":"10.1016\/j.carbpol.2010.04.025","article-title":"Modification of dextran\nusing click-chemistry approach in aqueous media","volume":"82","author":"Pahimanolis","year":"2010","journal-title":"Carbohydr. Polym."},{"key":"2026082516542544900_cit45","doi-asserted-by":"publisher","first-page":"939","DOI":"10.1038\/nm.3201","article-title":"A functional\nCFTR assay using primary cystic fibrosis intestinal organoids","volume":"19","author":"Dekkers","year":"2013","journal-title":"Nat. Med."},{"key":"2026082516542544900_cit46","doi-asserted-by":"publisher","first-page":"1500","DOI":"10.1002\/cbic.200700266","article-title":"Strong inhibition of cholera toxin by multivalent GM1\nderivatives","volume":"8","author":"Pukin","year":"2007","journal-title":"ChemBioChem"},{"key":"2026082516542544900_cit47","doi-asserted-by":"publisher","first-page":"117","DOI":"10.1016\/S0008-6215(00)82945-6","article-title":"The acid hydrolysis of substituted\nphenyl alpha-D-galactopyranosides","volume":"33","author":"De Bruyne","year":"1974","journal-title":"Carbohydr.\nRes."},{"key":"2026082516542544900_cit48","doi-asserted-by":"publisher","first-page":"4140","DOI":"10.1021\/acsnano.7b08479","article-title":"Quantitative Prediction of Multivalent Ligand-Receptor\nBinding Affinities for Influenza, Cholera, and Anthrax Inhibition","volume":"12","author":"Liese","year":"2018","journal-title":"ACS Nano"},{"key":"2026082516542544900_cit49","doi-asserted-by":"publisher","first-page":"471","DOI":"10.1002\/open.201500006","article-title":"Tetra- versus Pentavalent\nInhibitors of Cholera Toxin","volume":"4","author":"Fu","year":"2015","journal-title":"ChemistryOpen"},{"key":"2026082516542544900_cit50","doi-asserted-by":"publisher","first-page":"329","DOI":"10.1002\/cbic.200800550","article-title":"The Influence\nof Ligand Valency on Aggregation Mechanisms for Inhibiting Bacterial\nToxins","volume":"10","author":"Sisu","year":"2009","journal-title":"ChemBioChem"},{"key":"2026082516542544900_cit51","doi-asserted-by":"publisher","first-page":"12991","DOI":"10.1021\/ja027584k","article-title":"Solution and crystallographic\nstudies of branched multivalent ligands that inhibit the receptor-binding\nof cholera toxin","volume":"124","author":"Zhang","year":"2002","journal-title":"J. Am. Chem. Soc."},{"key":"2026082516542544900_cit52","doi-asserted-by":"publisher","first-page":"305","DOI":"10.1136\/oem.35.4.305","article-title":"The effect of acrylamide\non human polymorphonuclear\nneutrophils in vitro","volume":"35","author":"Mangan","year":"1978","journal-title":"Occup. Environ. Med."},{"key":"2026082516542544900_cit53","doi-asserted-by":"publisher","first-page":"419","DOI":"10.2147\/DDDT.S129922","article-title":"Conjugation\nof metronidazole with\ndextran: A potential pharmaceutical strategy to control colonic distribution\nof the anti-amebic drug susceptible to metabolism by colonic microbes","volume":"11","author":"Kim","year":"2017","journal-title":"Drug Des., Dev. Ther."},{"key":"2026082516542544900_cit54","doi-asserted-by":"publisher","first-page":"5931","DOI":"10.1002\/anie.201702005","article-title":"Multivalent Peptide?Nanoparticle\nConjugates for Influenza-Virus Inhibition","volume":"56","author":"Lauster","year":"2017","journal-title":"Angew.\nChem., Int. Ed."}],"container-title":["Bioconjugate Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.bioconjchem.8b00902","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/bcches\/article-pdf\/30\/3\/785\/67145394\/acs.bioconjchem.8b00902.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/bcches\/article-pdf\/30\/3\/785\/67145394\/acs.bioconjchem.8b00902.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T20:54:35Z","timestamp":1787691275000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/bcches\/article\/30\/3\/785\/586805\/Strong-Inhibition-of-Cholera-Toxin-B-Subunit-by"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,1,10]]},"references-count":55,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2019,1,10]]},"published-print":{"date-parts":[[2019,3,20]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.bioconjchem.8b00902","relation":{},"ISSN":["1043-1802","1520-4812"],"issn-type":[{"value":"1043-1802","type":"print"},{"value":"1520-4812","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,1,10]]}}}