In situ extension as an approach for identifying novel alpha-amylase inhibitors.
A new approach for the discovery and subsequent structural elucidation of oligosaccharide-based inhibitors of alpha-amylases based upon autoglucosylation of known alpha-glucosidase inhibitors is presented. This concept, highly analogous to what is hypothesized to occur with acarbose, is demonstrated with the known alpha-glucosidase inhibitor, d-gluconohydroximino-1,5-lactam. This was transformed from an inhibitor of human pancreatic alpha-amylase with a K(i) value of 18 mm to a trisaccharide analogue with a K(i) value of 25 mum. The three-dimensional structure of this complex was determined by x-ray crystallography and represents the first such structure determined with this class of inhibitors in any alpha-glycosidase. This approach to the discovery and structural analysis of amylase inhibitors should be generally applicable to other endoglucosidases and readily adaptable to a high throughput format.
Department of Chemistry, University of British Columbia, Vancouver, British Columbia V6T 1Z1, Canada.
Shin
S
Numao
Iben
I
Damager
Chunmin
C
Li
Tanja M
TM
Wrodnigg
Anjuman
A
Begum
Christopher M
CM
Overall
Gary D
GD
Brayer
Stephen G
SG
Withers
D020909:Acarbose / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D002240:Carbohydrate Sequence; D018360:Crystallography, X-Ray; D015195:Drug Design; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D008958:Models, Molecular; D015394:Molecular Structure; D010179:Pancreas / Q000201:enzymology; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D014312:Trisaccharides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D000516:alpha-Amylases / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000378:metabolism
D004791:Enzyme Inhibitors; D014312:Trisaccharides; D000516:alpha-Amylases; D020909:Acarbose