The goal of this Phase I proposal is to clone the gene for Amadorase, the enzyme responsible for the production of 3-deoxyglucosone (3DG). 3DG has been shown to be extremely important in the formation of protein cross-links and advanced glycation end products (AGEs). The formation of AGE products leads to diabetic complications such as nephropathy, retinopathy and neuropathy. An estimated 16 million people in the US suffer from type I or type II diabetes, with approximately 900,000 new cases diagnosed each year. Despite improvements in controlling blood glucose, current therapies are unable to prevent diabetic complications. These complications are associated with substantially increased health care requirements and a decreased life expectancy. Dynamis has discovered an enzymatically-mediated step in a lysine recovery pathway where a dinase, Amadorase, phosphorylates fructoselysine to produce the unstable intermediate fructoselysine 3-phosphate, that rapidly decomposes to yield 3DG. Because of its catalytic role in the production of 3DG, Amadorase is a promising new therapeutic target for the prevention of diabetic complications. Amadorase will be cloned by RT-PCR and the cloned protein tested enzymatically to demonstrate that it is Amadorase. The cloned gene will strongly facilitate the Phase II development of an HTS for potent Amadorase inhibitors. PROPOSED COMMERCIAL APPLICATIONS: Insulin dependent diabetes mellitus affects 0.5 to 1 million Americans while another 14 to 15 million people are affected by non-insulin dependent diabetes mellitus. Approximately 600,000 new cases of NIDDM and 30,000 new cases of IDDM are diagnosed each year. As well as developing other complications, forty percent of IDDM patients and 10 to 15 percent of NIDDM patients develop nephropathy. This amounts to approximately 1.6 to 2.7 million diabetics affected by nephropathy alone. Assuming $1-2000/person/year, the total sales potential for therapeutics able to ameliorate just diabetic nephropathy is in the range of $1-5 billion/year.
Thesaurus Terms: diabetes mellitus therapy, diabetic nephropathy, high throughput technology, molecular cloning, technology /technique development crosslink, deoxyglucose, diabetic neuropathy, diabetic retinopathy, enzyme activity, glycation, lysine, phosphorylation SDS polyacrylamide gel electrophoresis, fluorescence resonance energy transfer, laboratory rat, polymerase chain reaction, protein purification, tissue /cell culture
