Molecular glues for manipulating enzymes: trypsin inhibition by benzamidine-conjugated molecular glues

摘要

Water-soluble bioadhesive polymers bearing multiple guanidinium ion (Gu(+)) pendants at their side-chain termini (Glue(n)-BA, n = 10 and 29) that were conjugated with benzamidine (BA) as a trypsin inhibitor were developed. The Glue(n)-BA molecules are supposed to adhere to oxyanionic regions of the trypsin surface, even in buffer, via a multivalent Gu(+)/oxyanion salt-bridge interaction, such that their BA group properly blocks the substrate-binding site. In fact, Glue(10)-BA and Glue(29)-BA exhibited 35- and 200-fold higher affinities for trypsin, respectively, than a BA derivative without the glue moiety (TEG-BA). Most importantly, Glue(10)-BA inhibited the protease activity of trypsin 13-fold more than TEG-BA. In sharp contrast, (m)Glue(27)-BA, which bears 27 Gu(+) units along the main chain and has a 5-fold higher affinity than TEG-BA for trypsin, was inferior even to TEG-BA for trypsin inhibition.