title: Thrombolysis vs. bleeding from hemostatic sites by a prourokinase mutant compared with tissue plasminogen activator.
abstract: BACKGROUND
  A single site mutant (M5) of prourokinase (proUK) was developed to make proUK less vulnerable to spontaneous activation in plasma. This was a problem that seriously compromised proUK in clinical trials, as it precluded proUK-mediated fibrinolysis at therapeutic concentrations.
  
  
  RESULTS
  After completing dose-finding studies, 12 anesthetized dogs with femoral artery thrombosis were given either M5 (2.0 mg kg(-1)) or tissue plasminogen activator (t-PA) (1.4 mg kg(-1)) by i.v. infusion over 60 min (20% administered as a bolus). Two pairs of standardized injuries were inflicted at which hemostasis was completed prior to drug administration. Blood loss was quantified by measuring the hemoglobin in blood absorbed from these sites. Thrombolysis was evaluated at 90 min and was comparably effective by both activators. Rethrombosis developed in one t-PA dog. The principal difference found was that blood loss was 10-fold higher with t-PA (mean approximately 40 mL) than with M5 (mean approximately 4 mL) (P = 0.026) and occurred at more multiple sites (mean 2.7 vs. 1.2). This effect was postulated to be related to differences in the mechanism of plasminogen activation by t-PA and M5 in which the latter is promoted by degraded rather than intact (hemostatic) fibrin. In addition, two-chain M5 was efficiently inactivated by plasma C1 inactivator, an exceptional property which helped contain its non-specific proteolytic effect.
  
  
  CONCLUSIONS
  Intravascular thrombolysis by M5 was accompanied by significantly less bleeding from hemostatic sites than by t-PA. This was attributed to the proUK paradigm of fibrinolysis being retained at therapeutic concentrations by the mutation.
authors.affiliation: Vascular Research Laboratory, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA. vgurewic@bidmc.harvard.edu
authors.forename: V
authors.initials: V
authors.lastname: Gurewich
authors.forename: R
authors.initials: R
authors.lastname: Pannell
authors.forename: A
authors.initials: A
authors.lastname: Simmons-Byrd
authors.forename: P
authors.initials: P
authors.lastname: Sarmientos
authors.forename: J-N
authors.initials: JN
authors.lastname: Liu
authors.forename: S F
authors.initials: SF
authors.lastname: Badylak
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D004285:Dogs; D004795:Enzyme Stability / Q000235:genetics; D005263:Femoral Artery; D005342:Fibrinolysis / Q000187:drug effects / Q000235:genetics; D006470:Hemorrhage / Q000139:chemically induced*; D006487:Hemostasis / Q000187:drug effects*; D009154:Mutation; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000235:genetics; D015912:Thrombolytic Therapy / Q000009:adverse effects / Q000379:methods*; D013927:Thrombosis / Q000188:drug therapy; D010959:Tissue Plasminogen Activator / Q000008:administration & dosage*; D014568:Urokinase-Type Plasminogen Activator / Q000008:administration & dosage* / Q000235:genetics*
chemical_list: D011994:Recombinant Proteins; D010959:Tissue Plasminogen Activator; D014568:Urokinase-Type Plasminogen Activator; C067551:saruplase