Department of Chemical Biology and Drug Discovery, Utrecht Institute for Pharmaceutical Sciences , Utrecht University , Universiteitsweg 99 , 3584 CG Utrecht , The Netherlands.
Department of Pediatric Pulmonology, Regenerative Medicine Center Utrecht , University Medical Centre Utrecht , Lundlaan 6 , 3508 GA Utrecht , The Netherlands.
Bioconjug Chem. 2019 Mar 20;30(3):785-792. doi: 10.1021/acs.bioconjchem.8b00902. Epub 2019 Jan 24.
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 AB 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 α-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 IC values in the nanomolar range.
霍乱是一种潜在致命的细菌性感染，在发展中国家影响大量人群。它是由霍乱毒素（CT）引起的，霍乱毒素是由霍乱弧菌分泌的 AB 毒素。该毒素由一个毒性 A 亚单位和一个五聚体 B 亚单位组成，与肠道细胞表面结合。已经合成了几种单价和多价毒素抑制剂，但对于发展中国家的实际应用来说，它们过于复杂和昂贵。间硝基苯-α-半乳糖苷（MNPG）是 CT 的一种已知有前途的配体，这里合成了基于 MNPG 的单价和多价化合物。我们展示了 MNPG 的大大改进的产率及其在与多价支架连接时的用途。我们使用经济的聚合物作为多价支架，即聚丙烯酰胺、葡聚糖和超支化聚甘油（hPG）。铜催化的炔基叠氮化物环加成反应（CuAAC）产生了抑制剂，这些抑制剂在 ELISA 型测定和肠道类器官肿胀抑制测定中进行了测试。根据聚合物的类型，抑制特性差异很大，最有效的缀合物显示出纳摩尔范围内的 IC 值。
