title: Development of plasma kallikrein selective inhibitors.
abstract: During the course of the development of active center-directed plasmin inhibitors, it was found that N-(trans-4-aminomethylcyclohexanecarbonyl)-lysine-4-methoxycarb onylanilide 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-carboxy methyl- anilide (Tra-Phe-APAA). Tra-Phe-APAA inhibited plasma kallikrein with a Ki value of 0.81 microM, 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 microM, 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 glycinehydrochloric acid buffer (pH 3.0). Plasma kallikrein was purified 181-fold with a yield of 85% from the kaolin-activated plasma.
authors.affiliation: Department of Medicinal Chemistry, Faculty of Pharmaceutical Sciences, Kobe Gakuin University, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Okada
authors.forename: Y
authors.initials: Y
authors.lastname: Tsuda
authors.forename: M
authors.initials: M
authors.lastname: Tada
authors.forename: K
authors.initials: K
authors.lastname: Wanaka
authors.forename: A
authors.initials: A
authors.lastname: Hijikata-Okunomiya
authors.forename: U
authors.initials: U
authors.lastname: Okamoto
authors.forename: S
authors.initials: S
authors.lastname: Okamoto
mesh_terms: D001665:Binding Sites; D015195:Drug Design; D005341:Fibrinolysin / Q000037:antagonists & inhibitors; D006801:Humans; D007610:Kallikreins / Q000037:antagonists & inhibitors* / Q000737:chemistry; D007700:Kinetics; D015394:Molecular Structure; D009842:Oligopeptides / Q000737:chemistry / Q000494:pharmacology; D010649:Phenylalanine / Q000031:analogs & derivatives / Q000737:chemistry / Q000494:pharmacology; D015842:Serine Proteinase Inhibitors / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D013917:Thrombin / Q000037:antagonists & inhibitors; D014148:Tranexamic Acid / Q000031:analogs & derivatives / Q000737:chemistry / Q000494:pharmacology; D014568:Urokinase-Type Plasminogen Activator / Q000037:antagonists & inhibitors / Q000097:blood
chemical_list: D009842:Oligopeptides; D015842:Serine Proteinase Inhibitors; C064866:PKSI 527; D010649:Phenylalanine; C018788:valyl-leucyl-lysine 4-nitroanilide; D014148:Tranexamic Acid; D007610:Kallikreins; D013917:Thrombin; D005341:Fibrinolysin; D014568:Urokinase-Type Plasminogen Activator