İnsülin parçalayan enzimi regule eden moleküllerin biyokimyasal karakterizasyonu

Insulin-degrading enzyme (IDE) is an allosteric Zn+2 metalloprotease enzyme involved in the degradation of many peptides including amyloid beta (Aß), and insulin that play key roles in Alzheimer?s disease (AD) and type 2 diabetes mellitus (T2DM), respectively. Therefore, the regulation of IDE by small molecules is a rational approach for the treatment of these diseases by controlling the concentrations of substrates of enzyme. Crystal structure of IDE revealed that N-terminal of IDE has an exosite which is ~30 Å away from the catalytic region, and serves as a regulation site by orientation of the substrates of IDE to the catalytic site. Previous studies show that a 9-amino acid long peptide, bradykinin binds to the exosite and hence enhances the activity of IDE, which indicates that novel small molecules may bind to this exosite to regulate its activity. In this thesis, biochemical characterization of small molecules on the proteolytic activity of IDE was discovered using various physiological substrates of IDE. The effects of the molecules were monitored on the activity of IDE using different substrates. The resulting novel compounds, designated as D3, D4, and D6, enhances IDE mediated proteolysis of substrate V, insulin and FAßB, respectively. These selected compounds demonstrated submicromolar activation and can be utilized for the lead optimization studies to regulate substrate specific IDE activity. This study describes the first examples of a computer-aided discovery of IDE regulators, showing that in vitro activation of this important enzyme with drug-like small molecules is attainable.