Abiotic Mimic of Endogenous Tissue Inhibitors of Metalloproteinases: Engineering Synthetic Polymer Nanoparticles for Use as a Broad-Spectrum Metalloproteinase Inhibitor

Abstract

We describe a process for engineering a synthetic polymer nanoparticle (NP) that functions as an effective, broad-spectrum metalloproteinase inhibitor. Inhibition is achieved by incorporating three functional elements in the NP: a group that interacts with the catalytic zinc ion, functionality that enhances affinity to the substrate-binding pocket, and fine-tuning of the chemical composition of the polymer to strengthen NP affinity for the enzyme surface. The approach is validated by synthesis of a NP that sequesters and inhibits the proteolytic activity of snake venom metalloproteinases from five clinically relevant species of snakes. The mechanism of action of the NP mimics that of endogenous tissue inhibitors of metalloproteinases. The strategy provides a general design principle for synthesizing abiotic polymer inhibitors of enzymes.

A Biomimetic of Endogenous Tissue Inhibitors of Metalloproteinases: Inhibition Mechanism and Contribution of Composition, Polymer Size, and Shape to the Inhibitory Effect

“…5 nm, respectively (Figure A) . In our previous study of hydrogel nanoparticle (NP) inhibitors of whole venom, the two functional monomers (6-methacrylamidohexanoyl)- S -phenylalanine hydroxamate (PHX), a binding group to the catalytic zinc cation and substrate pocket, and/or N -acrylamido- S -phenylalanine (APhe), a binding group to enhance polymer affinity to the enzyme surface, were found to be important for inhibition. In the present study these monomers were among those included in N -isopropylacrylamide (NIPAm)-based linear copolymers using controlled radical polymerization and reversible addition–fragmentation chain transfer (RAFT) polymerization (Figure B).…”

“…Multipoint interactions are a design principle for synthetic polymer/nanoparticles (NPs) engineered with an affinity for target biomacromolecules. − Their high molecular weight enables coverage of significant portions of a protein surface. Polymer/NP ligands also often function as enzyme inhibitors by binding to the vicinity of the active site of the target enzyme. − For example, poly- N -isopropylacrylamide (NIPAm)-based NP copolymers optimized to inhibit enzymes from the venoms of multiple species of snakes are of interest as abiotic antidotes with potential use in the treatment of snakebite envenoming. , We recently reported a polymer NP that mimics the mechanism of the action of TIMPs by optimizing the combination/population of binding elements for the catalytic zinc cation, the substrate-binding pocket, and the enzyme surface …”

Nanobiotechnology with Therapeutically Relevant Macromolecules from Animal Venoms: Venoms, Toxins, and Antimicrobial Peptides

“…Using synthetic PNs, other antivenoms were developed from other snakes, such as Crotalus atrox , Bitis arietans , Bitis gabonica , Echis ocellatus , and Echis carinatus. The inactivation of metalloproteinases could be achieved for all of the species studied, which is interesting for the development of new enzyme inhibitors that are effective against snake bites [ 56 ]. The authors recommended caution when trying to obtain nanoparticles of protein or peptide bioactive compounds, as some methods/time of preparation or some compounds of the preparation solutions could break or inactivate a peptide bond.…”

Biomolecular interactions of ultrasmall metallic nanoparticles and nanoclusters

“…[64][65][66] Of note, large NP scaffolds with tunable surface chemistries have also been under development for protein surface recognition and the control of protein binding orientation. [67][68][69] However, despite successful examples in this area, realizing specic protein surface recognition with ultrasmall or larger NPs alike remains an enormous challenge. The reason is that specic recognition requires proper chemical and surface complementarity at the NP/protein binding interface, an accepted notion in highaffinity protein-protein binding and more apparent in usNPprotein associations.…”