CHARACTERIZATION OF A CLASS OF PEPTIDE BORONATES WITH NEUTRAL P1 SIDE-CHAINS AS HIGHLY SELECTIVE INHIBITORS OF THROMBIN

Z-D-Phe-Pro-boroMpg-OPin (9a)(1,2) has been shown previously to be a highly specific inhibitor of thrombin in spite of lacking an arginine-like guanidino group at the P1 site.(1) A range of compounds have been synthesized based upon this lead compound, varying the neutral side chain at the P1 site. Of the 20 examples based upon the structures at P2 and P3 of Z-D-X-Pro (X being Phe or beta,beta-diphenylalanine), all were found to be effective inhibitors of thrombin (K-i's between 10 and 100 nM). Furthermore all exhibited a high specificity toward thrombin having values for st K-i(trypsin)/K-i(thrombin) ratio of between 10- and 100-fold. High ratio values were found for a number of the compounds tested against a range of serine proteinases (plasmin, factor Xa, kallikrein, urokinase, protein Ca, chymotrypsin, elastase, and cathepsin G).(2) AS far as potency toward thrombin, compounds containing the methoxypropyl group at P1 were favored over those with a methoxy grouping on a shorter alkyl chain (8) or without the methoxy group (1-5). The compounds display potent anticoagulant activity with values for 18 in thrombin time of 0.63 mu M and in activated partial thromboplastin time of 2.0 mu M. B-11 NMR has been used to confirm interaction of the boron atom with the active site. From the high specificity shown with all the compounds we propose that the compounds, constitute a new class of thrombin inhibitors.