With aim to deliver vegetal copper-containing amine oxidase (Cu-AO) to the intestine in the presence of catalase as a novel way to control intestinal dysfunctions connected with histamine-related conditions such as allergic/pseudoallergic reactions, colon cancer, Crohn's disease, ulcerative colite, the project here presented will be focus either to gain a better understanding of the mechanisms of the interaction of vegetal Cu-AO, an efficient histaminases, alone or in combination with catalase, with cells or to assess the stability of the enzyme in the intestinal environment. To achieve these goals, the project will be performed in collaboration with Prof. Mircea Alexandru Mateescu of the University of Quebec at Montreal and will be articulated in the following parts:
1) purification of Cu-AO from Lathyrus sativus seedlings and improvement of the procedures to prepare extracts containing Cu-AO and catalase at concentrations appropriate to remove efficiently both histamine and hydrogen peroxide;
2) evaluation of ability of Cu-AO alone or in combination with catalase and histamine to bind to a human colon colorectal adenoma derived cells (CaCo2) or to rat intestinal tissue;
3) analysis of the effect of vegetal Cu-AO alone or in combination with catalase and histamine on the calcium homeostasis in CaCo2 cells;
4) assessment of the stability of Cu-AO and catalase in simulated intestinal fluids.
The study will provide new data to prepare formulations for oral supplementation able to target and deliver Cu-AO and catalase to the intestinal sites of inflammation in a catalytic active forms. The gastric protection against stomach acidity of the enzymes and their controlled release to the colon is the key to treatment and extension to novel forms of interest for nanotechnology, such as nanoparticles, microspheres and tablets will be made. Collaborations with Pharmaqam and BioMed groups (Montreal, Canada) and Galenica Senese (Siena, Italy) will be envisaged.
