EnzyMime - Biocompatible and sustainable enzyme mimics based on metal complexes with peptide derivatives: synthesis, characterization and potential biological applications
Abstract:
The development of efficient EMs represents a formidable opportunity on the way of a green transition and a high grade of competitive innovation, both needful for our prosperity and wellbeing. Notably, in recent years, the research progress on EMs has led to significant developments of sustainable chemical synthesis protocols, at both laboratory and industrial scale level. These compounds have the potential to compete with natural enzymes and “classical” catalysts, finding application in a wide range of fields, from chemical and food industries to diagnostics and medicine or production of fuels and gaseous hydrogen.
EMs are usually designed to reproduce the catalytic sites of natural enzymes, following two possible approaches: imitating the enzyme functionality using metal complexes with similar activity, or mimicking the structure of the enzyme active site by introducing appropriate functional groups (e.g.: oligopeptides).
The present research project (EnzyMime) aims at designing a new class of de novo EMs, based on synthetic branched peptides. A biocompatible central scaffold represents the core of the EM structure, to which four identical oligopeptides are linked through the so-called “peptide welding technology” (PWT). This will allow the easy development of different EMs, where the potential catalytic site can be produced introducing appropriate amino acid sequences, able to bind the active metal ions. An example may be represented by the synthesis of different surrogates of Cu/Zn-SOD, Mn-SOD and Ni-SOD to catalyse dismutation of superoxide radicals.
The experimental approach will consist of: synthesis of several EMs and their catalytic characterization; complete characterization of the most promising metal/peptide systems, through potentiometry, voltammetry, spectrophotometry, spectrofluorimetry, mass spectrometry, nuclear magnetic resonance, electron paramagnetic resonance, theoretical calculations and structural investigation by X-ray crystallography. To help the crystallization of peptides and their metal complexes, a spacer containing a short alpha-helix folding sequence will also been added to the chelating sequences. Finally, in order to explore some possible practical applications in the field of human health and wellbeing, biological tests will be performed, including enzymatic stability in human plasma, interaction with nucleic acids, cytotoxicity against cancer cells, mobility through different types of membranes and antimicrobial activity.
Risultati attesi:
The ad hoc design of enzyme mimics (EMs) based on branched peptides and with specific and effective metal binding sites can be a successful approach to obtain novel bioactive molecules characterized by an extraordinary versatility.
The synthesis and characterization of effective EMs will promote and strengthen scientific research in the biomedical area for the following reasons. The therapeutic use of our metal-EMs with antioxidant scope can improve the health conditions of citizens suffering from several chronic and degenerative diseases. What is more outstanding, however, is the potential substrate versatility of EMs which make them useful for many other applications. The results of this project will give important information on the capacity of these systems to perform not only an antioxidant activity but also to intervene in other cellular redox processes. For instance, the evaluation of ROS production and interaction with DNA will allow us to consider the use of our EMs as antiproliferative agents against cancer.
Tetrabranched peptides can also be used for imaging applications, relying on their higher metal-binding stability to form more stable complexes or increasing the local concentration of radioactive isotopes per molecule.
It is also worth of note that the redox catalytic activity can be useful for other applications different from the biomedical ones, as, for example, for environmental decontamination, including degradation of organic pollutants and chelation of heavy metal ions in soils and waters.
Dettagli progetto:
Referente scientifico: Bellotti Denise
Fonte di finanziamento: Bando PRIN 2022 PNRR
Data di avvio: 30/11/2023
Data di fine: 30/11/2025
Contributo MUR: 60.200 €
Partner:
- Università degli Studi di FERRARA (capofila)
- Università degli Studi di PALERMO
- Università degli Studi di SASSARI
- Università degli Studi di Napoli Federico II
- Università degli Studi di TRIESTE
- Università degli Studi di TORINO