Inhibition of aspartic proteinases by peptides containing lysine and ornithine side-chain analogues of statine.
The synthesis of two new analogues of statine are reported corresponding to analogues with the lysine side chain and the ornithine side chain. These analogues were designed on the basis of substrate specificity and molecular modeling of three-dimensional structures of the penicillopepsin: Iva-Val-Sta-OEt crystal structure. 4,8-Diamino-3-hydroxyoctanoic acid [LySta] and 4,7-diamino-3-hydroxyheptanoic acid [OrnSta] were synthesized respectively from Boc-Lys(Z)-al and Boc-Orn(Bzl,Z)-al by addition of lithio ethyl acetate to the aldehyde group. The [LySta] derivative was converted to the trichloroethoxycarbonyl derivative and separated into the corresponding 3S,4S and 3R,4R diastereomers. The [OrnSta] derivative was used as a mixture of 3-position diastereomers. These new amino acids were used to prepare the following inhibitors: Iva-Val-Val-[LySta]-OEt and Iva-Val-Val-[OrnSta]-OEt as well as the corresponding synthetic intermediates. Inhibition constants (Ki values) were measured for inhibition of porcine pepsin and penicillopepsin. Both compounds were potent inhibitors of penicillopepsin with Ki values 10-100 times smaller (2.1 and 1.1 nM, respectively) than the Ki of Iva-Val-Val-Sta-OEt (47 nM). In contrast both inhibitors are exceptionally weak inhibitors of porcine pepsin with Ki values greater than 1 microM. These results are correlated with the ability of the basic group in the new inhibitors to bind to aspartic acid-77 in penicillopepsin.
D000596:Amino Acids; D016282:Aspartic Acid Endopeptidases; D010450:Endopeptidases; D007202:Indicators and Reagents; D007700:Kinetics; D008239:Lysine; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D009952:Ornithine; D010455:Peptides / Q000138:chemical synthesis* / Q000494:pharmacology; D011480:Protease Inhibitors / Q000138:chemical synthesis*; D011487:Protein Conformation; D013329:Structure-Activity Relationship
D000596:Amino Acids; D007202:Indicators and Reagents; D010455:Peptides; D011480:Protease Inhibitors; D009952:Ornithine; D010450:Endopeptidases; D016282:Aspartic Acid Endopeptidases; D008239:Lysine; C013165:statine