CX-397, a novel recombinant hirudin analog having a hybrid sequence of hirudin variants-1 and -3.
To elucidate the differential roles of N- and C-terminal halves of hirudins in thrombin inhibition, we produced novel recombinant hirudin analogs, CX-397 and CX-397R, having a hybrid amino acid sequence of hirudin variants-1 (HV-1) and -3 (HV-3). CX-397 is composed of the N-terminal half of HV-1 (HV-11-36) combined with the C-terminal half of HV-3 (HV-337-66). CX-397R is the opposite combination. Their anti-thrombin activity was determined by a fluorogenic enzyme assay and compared with that of recombinant HV-1 (rHV-1) and rHV-3. The order of the magnitude of dissociation constants (Ki) of these four hirudin analogs in thrombin inhibition was as follows: CX-397R (0.294 pM) > rHV-1 (0.148 pM) > rHV-3 (0.0593 pM) > CX-397 (0.0433 pM), indicating that CX-397 is the strongest inhibitor among them.
Komatsu
Y
Y
Pharmaceuticals & Biotechnology Laboratory, Nikko Kyodo Co., Ltd., Saitama, Japan.
Misawa
S
S
Sukesada
A
A
Ohba
Y
Y
Hayashi
H
H
D000595:Amino Acid Sequence; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli; D006629:Hirudins / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000302:isolation & purification / Q000494:pharmacology*; D006801:Humans; D007700:Kinetics; D008433:Mathematics; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D013917:Thrombin / Q000037:antagonists & inhibitors* / Q000302:isolation & purification
D006629:Hirudins; D011994:Recombinant Proteins; D013917:Thrombin