Identification of heparanase-recognizing HS sequences. Affinity chromatography on immobilised heparanase. Sequencing of radiolabeled HS oligosaccharides with high affinity for the enzyme. Molecular modeling, NMR spectroscopic, and X-ray diffraction studies on recombinant heparanase. Binding studies in solution with polysaccharide fragments and with synthetic oligosaccharides, by affinity chromatography, electrophoresis, MALDI and NMR spectroscopy. Crystallization and X-ray studies performed on heparanase and on any crystallizable oligosaccharide-heparanase complexes.
Generation of heparanase-inhibiting, antiangiogenic and antimetastatic substances. Starting form preparation and structural characterization of sulfated polysaccharides (such as laminaran sulfate) and oligosaccharides (maltohexaose sulfate) already established as heparanase inhibitors, synthesis of novel potential inhibitors obtained by regioselective sulfation of other polyglucoses and bacterial glycosaminoglycans, as well as glucosidase-resistant glycomimetic oligosaccharides. Testing products for growth inhibition of tumors produced by heparanase-secreting cells. Selection of lead compounds.