On-bead combinatorial techniques for the identification of selective aldose reductase inhibitors.
Aldose reductase (AKR1B1; ALR2; E.C. 1.1.1.21) is an NADPH-dependent carbonyl reductase which has long been associated with complications resulting from the elevated blood glucose often found in diabetics. The development of effective inhibitors has been plagued by lack of specificity which has led to side effects in clinical trials. To address this problem, a library of bead-immobilized compounds was screened against fluorescently labeled aldose reductase in the presence of fluorescently labeled aldehyde reductase, a non-target enzyme, to identify compounds which were aldose reductase specific. Picked beads were decoded via novel bifunctional bead mass spec-based techniques and kinetic analysis of the ten inhibitors which were identified using this protocol yielded IC50 values in the micromolar range. Most importantly, all of these compounds showed a preference for aldose reductase with selectivities as high as approximately 7500-fold. The most potent of these exhibited uncompetitive inhibition versus the carbonyl-containing substrate D/L-glyceraldehyde with a Ki of 1.16 microM.
Robins
Lori I
LI
Department of Chemistry, University of California, One Shields Avenue, Davis, CA 95616, USA.
Dixon
Seth M
SM
Wilson
David K
DK
Kurth
Mark J
MJ
D000449:Aldehyde Reductase / Q000037:antagonists & inhibitors*; D020650:Combinatorial Chemistry Techniques / Q000379:methods*; D048909:Diabetes Complications / Q000188:drug therapy; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004791:Enzyme Inhibitors / Q000302:isolation & purification; D006801:Humans; D020128:Inhibitory Concentration 50; D008863:Microspheres; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
D004791:Enzyme Inhibitors; D000449:Aldehyde Reductase