S2′‐subsite variations between human and mouse enzymes (plasmin, factor XIa, kallikrein) elucidate inhibition differences by tissue factor pathway inhibitor ‐2 domain1‐wild‐type, Leu17Arg‐mutant and aprotinin. (19th November 2016)

• Record Type: Journal Article
• Authors: Vadivel, K.; Kumar, Y.; Ogueli, G. I.; Ponnuraj, S. M.; Wongkongkathep, P.; Loo, J. A.; Bajaj, M. S.; Bajaj, S. P.
• Is Part Of: Journal of thrombosis and haemostasis. Volume 14:Number 12(2016:Dec.)
• Page Start: 2509
• Page End: 2523
• Publication Date: 2016-11-19
• Subjects: antifibrinolytic agents -- aprotinin -- fibrinolysis -- TFPI‐2 -- tranexamic acid; Thrombosis -- Periodicals; Hemostasis -- Periodicals; Blood coagulation disorders -- Periodicals; 616.1
• DOI: 10.1111/jth.13538
• Languages: English
• ISSNs: 1538-7933

Abstract

Abstract : Essentials Current antifibrinolytics – aminocaproic acid and tranexamic acid–can cause seizures or renal injury. KD1L17R ‐KT, aprotinin and tranexamic acid were tested in a modified mouse tail‐amputation model. S2′‐subsite variations between human and mouse factor XIa result in vastly different inhibition profiles. KD1L17R ‐KT reduces blood loss and D‐dimer levels in mouse with unobserved seizures or renal injury. Summary: Background: Using tissue factor pathway inhibitor (TFPI)‐2 Kunitz domain1 (KD1), we obtained a bifunctional antifibrinolytic molecule (KD1L17R ‐KT ) with C‐terminal lysine (kringle domain binding) and P2′‐residue arginine (improved specificity towards plasmin). KD1L17R ‐KT strongly inhibited human plasmin (hPm), with no inhibition of human kallikrein (hKLK) or factor XIa (hXIa). Furthermore, KD1L17R ‐KT reduced blood loss comparable to aprotinin in a mouse liver‐laceration model of organ hemorrhage. However, effectiveness of these antifibrinolytic agents in a model of hemorrhage mimicking extremity trauma and their inhibition efficiencies for mouse enzymes (mPm, mKLK or mXIa) remain to be determined. Objective: To determine potential differences in inhibition constants of various antifibrinolytic agents against mouse and human enzymes and test their effectiveness in a modified mouse tail‐amputation hemorrhage model. Methods/Results: Unexpectedly, mXIa was inhibited with ~ 17‐fold increased affinity by aprotinin ( K i ~ 20 nm ) and with measurable affinity for KD1L17R ‐KT ( K i ~ 3 μm ); in contrast, KD1WT ‐VT inhibited hXIa or mXIa with similar affinity. Compared with hPm, mPm had ~ 3‐fold reduced affinity, whereas species specificity for hKLK and mKLK was comparable for each inhibitor. S2′‐subsite variations largely accounted for the observed differences. KD1L17R ‐KT and aprotinin were more effective than KD1WT ‐VT or tranexamic acid in inhibiting tPA‐induced mouse plasma clot lysis. Further, KD1L17R ‐KT was more effective than KD1WT ‐VT and was comparable to aprotinin and tranexamic acid in reducing blood loss andD ‐dimer levels in the mouse tail‐amputation model. Conclusions: Inhibitor potencies differ between antifibrinolytic agents against human and mouse enzymes. KD1L17R ‐KT is effective in reducing blood loss in a tail‐amputation model that mimics extremity injury.