Title
PEG-DAAO conjugate: A promising tool for cancer therapy optimized by protein engineering
Author
Elena Rosini, Loredano Pollegioni
Year
2020
Journal
Nanomedicine: Nanotechnology, Biology and Medicine
Abstract
The flavoenzyme D-amino acid oxidase (DAAO) represents a potentially good option for cancer enzyme prodrug therapy as it produces H2O2 using D-amino acids as substrates, compounds present at low concentration in vivo and that can be safely administered to regulate H2O2 production. We optimized the cytotoxicity of the treatment by: i) using an efficient enzyme variant active at low O2 and D-alanine concentrations (mDAAO); ii) improving the stability and half-life of mDAAO and the enhanced permeability and retention effect by PEGylation; and iii) inhibiting the antioxidant cellular system by a heme oxygenase-1 inhibitor (ZnPP). A very low amount of PEG-mDAAO (10 mU, 50 ng of enzyme) induces cytotoxicity on various tumor cell lines. Notably, PEG-mDAAO seems well suited for in vivo evaluation as it shows the same cytotoxicity at air saturation (21%) and 2.5% O2, a condition resembling the microenvironment found in the central part of tumors.
Instrument
J-810, V-560, FP-750
Keywords
Absorption, Ligand binding, Fluorescence, Protein structure, Protein stability, Kinetics, Circular dichroism, Secondary structure, Thermal stability, Thermodynamics, Biochemistry, Materials, Medicinal