bm7b00162_si_001.pdf (5.44 MB)
High-Affinity Copolymers Inhibit Digestive Enzymes by Surface Recognition
journal contribution
posted on 2017-04-18, 00:00 authored by Patrick Gilles, Kirstin Wenck, Inga Stratmann, Michael Kirsch, Daniel A. Smolin, Torsten Schaller, Herbert de Groot, Arno Kraft, Thomas SchraderThis
account presents a general method for the construction of
polymeric surface binders for digestion enzymes. Two prominent parts,
namely, the modification of the copolymer composition and the screening
assay for the most powerful inhibitors are both amenable to parallelization.
The concept hinges on the appropriate selection of amino-acid-selective
comonomers, their free radical copolymerization, and subsequent screening
of the resulting copolymer library for efficient enzyme inhibition.
A microscale synthetic procedure for the copolymerization process
was developed, which produces water-soluble affinity polymers that
can be stored for years at room temperature. Initial parallel screening
was conducted in standard enzyme assays to identify polymeric inhibitors,
which were subsequently subjected to determination of IC50 values for their target enzyme. For all digestion enzymes, except
elastase, a number of polymer inhibitors were found, some of which
were selective toward one or two protein targets. Since the key monomers
of the best inhibitors bind to amino acid residues in the direct vicinity
of the active site, we conclude that efficient coverage of the immediate
environment by the copolymers is critical. Strong interference with
enzymatic activity is brought about by blocking the substrate access
and product exit to and from the active site.