A
mutant prourokinase
plasminogen activator (M5) was developed to make prouPA less subject to spontaneous conversion to tcuPA in blood at therapeutic concentrations. Two-chain M5 was shown to form complexes with C1-inhibitor, which was the principal inhibitor of tcM5 in
plasma. The effect of supplemental additions of C1-inhibitor on
fibrinolysis and fibrinogenolysis by M5 was determined. Supplemental C1-inhibitor restored the stability of high-
dose M5 and prevented fibrinogenolysis but not
fibrinolysis, the rate of which was not compromised by the inhibitor. Due to higher
dose tolerance of M5 in the presence of supplemental C1-inhibitor, the rate of
fibrin-
specific lysis reached that achievable by nonspecific
fibrinolysis, which is the maximum possible for a
plasminogen activator.
Plasma C1-inhibitor stabilized M5 in
plasma by inhibiting tcM5 which would otherwise greatly amplify non-specific plasminogen activation causing more tcM5 generation from M5. This unusual dissociation of inhibitory effects, whereby fibrinogenolysis and not fibrinolysis is inhibited, has significant implications for improving the safety and
efficacy of fibrinolysis. Methods of reducing bleeding and non-specific plasminogen activation during fibrinolysis by administering M5 along with exogenous C1-inhibitor are disclosed.