Title: Biomimetic materials for enzyme recognition and inhibition obtained by molecular imprinting.
Author: Arnaud CUTIVET
National thesis number: 2008COMP1776
- The aim of this thesis is the development of a new class of enzyme inhibitors exhibiting strong affinity and selectivity for a target enzyme, using molecular imprinting technology. Molecular imprinting allows to create complementary images or molecular imprints of a template molecule in a crosslinked synthetic polymer. In the present work, a new approach was developped where the polymerisation was conducted from the enzyme’s active site, using an anchoring compound from which the polymer chains grow in the proximity of the enzyme to form polymer microgels. As a model, imprinting of the protease trypsin was first studied; later, the technique was applied to glucosidases. The imprinted materials were prepared from a highly diluted aqueous solution. For trypsin imprinting, methacrylamido benzamidine was synthesized and used as anchoring monomer. The presence of molecular imprints was confirmed by binding assays, which showed that the imprinted polymers displayed a much higher affinity towards the target enzyme than the non-imprinted control polymers, as well as a clear selectivity for the enzyme over other proteins. When the inhibition properties were evaluated, trypsin-imprinted polymers showed stronger inhibition than the corresponding control polymers and the small inhibitor benzamidine. As an alternative way to confine microgel synthesis, an iniferter-based dendrimeric multi-initiator was employed. This molecule affords a high local radical concentration. Monodispersed spherical particles were obtained, exhibiting a more compact morphology than with conventional initiators. The use of these biomimetic materials as drugs is one of the potential applications.