HIGH THROUGHPUT SCREEN FOR AMADORASE INHIBITORS

DESCRIPTION (provided by applicant): The goal of this Phase I SBIR proposal is to develop a cell-based high throughput screen for inhibitors of a novel fructosamine 3-kinase present in the kidney. Dynamis has named this enzyme Amadorase since it phosphorylates fructoselysine, an Amadori product, to produce fructoselysine 3-phosphate (FL3P). FL3P is unstable and decomposes to lysine, inorganic phosphate, and 3-deoxyglucosone (3DG), a highly reactive dicarbonyl sugar. 3DG is a precursor to advanced glycation end products (AGEs), which play a role in the development of diabetic nephropathy, retinopathy, neuropathy, and heart disease. Since Amadorase is responsible for most of the in vivo production of 3DG, it is an exciting new potential therapeutic target for diabetic complications. Dynamis hypothesizes that the reduction of systemic 3DG levels will result in slowing of the development or a reduction in the severity of diabetic complications. Since there are presently no treatments to halt the progression of diabetic complications, any level of reduction obtained is extremely important to the health and well being of diabetics. Dynamis proposes to create a cell-based assay where growth of Amadorase-expressing, lysine auxotrophs of E. coli or yeast is dependent upon Amadorase regenerating lysine from fructoselysine. Inhibitors of Amadorase will thus inhibit cell growth. A scintillation proximity assay (SPA) is proposed as a fallback system. The HTS will be used in a Phase II SBIR to screen combinatorial chemistry libraries for new Amadorase inhibitors. The discovery of potent drug candidates will lead to strategic partnerships with companies capable of conducting Phase I-Ill clinical trials.