Development of plasma kallikrein selective inhibitors

• Published in: BIOPOLYM 1999, Vol.51 (1), p.41-50
• Main Authors: Okada, Yoshio, Tsuda, Yuko, Tada, Mayako, Wanaka, Keiko, Hijikata-Okunomiya, Akiko, Okamoto, Utako, Okamoto, Shosuke
• Format: Article
• Language: English
• Subjects: Adsorption; Binding Sites; Biosynthesis; Catalyst activity; Catalyst selectivity; Drug Design; Fibrinolysin - antagonists & inhibitors; Humans; Kallikreins - antagonists & inhibitors; Kallikreins - chemistry; Kinetics; Molecular Structure; N-(trans-4-aminomethylcyclohexanecarbonyl)-phenylalanine-4-carboxymethylanilide (Tra-Phe-APAA); new affinity gel; Oligopeptides - chemistry; Oligopeptides - pharmacology; Phenylalanine - analogs & derivatives; Phenylalanine - chemistry; Phenylalanine - pharmacology; PKSI-Toyopearl; plasma kallikrein selective inhibitors; Serine Proteinase Inhibitors - chemical synthesis; Serine Proteinase Inhibitors - chemistry; Serine Proteinase Inhibitors - pharmacology; Thrombin - antagonists & inhibitors; Tranexamic Acid - analogs & derivatives; Tranexamic Acid - chemistry; Tranexamic Acid - pharmacology; Urokinase-Type Plasminogen Activator - antagonists & inhibitors; Urokinase-Type Plasminogen Activator - blood
• ISSN: 0006-3525; 1097-0282
• DOI: 10.1002/(SICI)1097-0282(1999)51:1<41::AID-BIP5>3.0.CO;2-Y

During the course of the development of active center‐directed plasmin inhibitors, it was found that N‐(trans‐4‐aminomethylcyclohexanecarbonyl)‐lysine‐4‐methoxycarbonylanilide inhibited plasma kallikrein more potently than other enzymes such as plasmin, urokinase, and thrombin, although the inhibitory activity was not as potent and enzyme selectivity not as high. Based on studies of structure–activity relationship, we designed and synthesized the plasma kallikrein selective inhibitor, N‐(trans‐4‐aminomethylcyclohexanecarbonyl)‐phenylalanine‐4‐carboxymethylanilide (Tra‐Phe‐APAA). Tra‐Phe‐APAA inhibited plasma kallikrein with a Ki value of 0.81 μM, while it inhibited glandular kallikrein, plasmin, urokinase, tissue plasminogen activator, factor Xa, factor XIIa, and thrombin with Ki values of > 500, 390, 200, > 500, > 500, > 500, and > 500 μM, respectively. We designated Tra‐Phe‐APAA as PKSI‐527. Using PKSI‐527 as an affinity ligand, we synthesized a new affinity gel (PKSI‐Toyopearl) and employed it for the rapid purification of plasma kallikrein from human plasma. Human plasma activated with kaolin after acid treatment was applied to a PKSI‐527‐Toyopearl column. Adsorbed protein was eluted with 50 mM glycine‐hydrochloric acid buffer (pH 3.0). Plasma kallikrein was purified 181‐fold with a yield of 85% from the kaolin‐activated plasma. © 1999 John Wiley & Sons, Inc. Biopoly 51: 41–50, 1999