title: New bivalent thrombin inhibitors with N(alpha)(methyl)arginine at the P1-position.
abstract: A series of bivalent thrombin inhibitors was synthesized, consisting of a d-phenylalanyl-prolyl-N(alpha)(methyl)arginyl active site blocking segment, a fibrinogen recognition exosite inhibitor part, and a peptidic linker connecting these fragments. The methylation of the P1 amino acid led to a moderate decrease in affinity compared with the unmethylated analog. In addition, it prevented the thrombin catalyzed proteolysis, independent of the P1' amino acid used. This is a significant advantage compared to the original hirulogs, which strictly require a proline as P1' amino acid to reduce the cleavage C-terminal to the arginyl residue. Several analogs were prepared by incorporation of different P1' amino acids found in natural thrombin substrates. The most potent inhibitor was I-11 [dCha-Pro-N(Me)Arg-Thr-(Gly)5-DYEPIPEEA-Cha-dGlu] with a Ki of 37 pM. I-11 is highly selective and no inhibition of the related serine proteases trypsin, factor Xa and plasmin was observed. The stability of I-11 in human plasma in vitro was strongly improved compared to hirulog-1. In addition, a significantly reduced plasma clearance of I-11 was observed after intravenous injection in rats. Results from molecular modeling suggest that a strong reorganization of the hydrogen bonds in the active site of thrombin may result in the proteolytic stability found in this inhibitor series.
authors.affiliation: Institut für Biochemie und Biophysik, Friedrich-Schiller-Universität, Jena, Germany.
authors.forename: T
authors.initials: T
authors.lastname: Steinmetzer
authors.forename: M
authors.initials: M
authors.lastname: Batdordshjin
authors.forename: F
authors.initials: F
authors.lastname: Pineda
authors.forename: L
authors.initials: L
authors.lastname: Seyfarth
authors.forename: A
authors.initials: A
authors.lastname: Vogel
authors.forename: S
authors.initials: S
authors.lastname: Reissmann
authors.forename: J
authors.initials: J
authors.lastname: Hauptmann
authors.forename: J
authors.initials: J
authors.lastname: Stürzebecher
mesh_terms: D000595:Amino Acid Sequence; D000991:Antithrombins / Q000737:chemistry*; D006801:Humans; D008969:Molecular Sequence Data; D019323:omega-N-Methylarginine / Q000737:chemistry*
chemical_list: D000991:Antithrombins; D019323:omega-N-Methylarginine