Inhibition of platelets aggregation by [D-Phe(Transcinnamoyl)-Pro-D-Arg-P1'-CONH2] peptides reversible inhibitors of thrombin

BIOL 126

Cristina Clement, cclement_us@yahoo.com1, Anna Babinska, Anna.Babinska@downstate.edu2, Elizabeth Kornecki2, and Manfred Philipp, hwm_philipp@yahoo.com3. (1) Chemistry Department, Lehman College, City University of New York, 250 Bedford Park BLVD West, Bronx, New York City, NY 10468, (2) Department Anatomy and Cell Biology, SUNY, Down State Medical Center, 450 Clarkson Ave, Brooklyn-NYC, NY 11203, (3) Chemistry Department, Lehman College and Biochemistry Ph.D. Program, City University of New York, 365 Fifth Avenue, New York City, NY 10016-4309
A structure-activity relationship (SAR) for reversible inhibitory activity toward thrombin of tetrapetides from series D-Phe-Pro-DArg-P1'-CONH2 is reported. The P1' position was varied with D and L amino acids. The significant differences between the inhibitory constants (Kis) of tetrapeptides from the series D-Phe-Pro-D-Arg-P1'-CONH2 suggest that the interaction between the amino acid at P1' position and the S1' subpocket in thrombin is very specific. There is a 2 to 500 fold experimentally determined difference between the Kis of different peptide inhibitors and our in vitro inhibition assay for thrombin proved that the P1' position requires small and polar amino-acids. Functional assays, such as thrombin-induced platelets aggregation confirmed that the peptides were effective in inhibiting the platelets aggregation completely at concentration 10-15 fold their Ki determined from in vitro inhibition of thrombin. The aggregation of platelets by some lead peptides was specific only for thrombin-induced platelets aggregation, since there was no inhibition of platelets aggregation in the presence of another agonist for activation of such as collagen. These results strongly support our original structure-based design of peptides as reversible inhibitors for thrombin.