Pharmaceutical compositions of fibrinolytic agent
Frozen and lyophilized compositions for a metalloproteinase fibrinolytic agent (fibrolase or NAT), a method for preparing the lyophilized composition, and a kit and method for reconstituting the lyophilized composition are described herein.
1. A method for preparing a pharmaceutical composition comprising:
(a) providing an aqueous composition comprising a therapeutically effective amount of a fibrinolytic metalloproteinase, a zinc stabilizer and, optionally, a bulking agent, in a pharmaceutically acceptable buffer comprising citric acid or a water soluble citric acid salt;
(b) lyophilizing said aqueous composition; and
(c) reconstituing said lyophilized composition.
2. The method of claim 1 , wherein the aqueous composition comprises about 0.1 to about 50 mg/ml of the metalloproteinase, about 0.08 to about 0.12 mM of zinc sulfate, about 18 to about 22 mM of citric acid or sodium citrate, about 3 to about 6 mM of Tris, about 3 to about 6 percent (w/v) of mannitol, and about 0.008 to about 0.012 percent (w/v) of a surfactant, and optionally about 0.1 to about 0.5 percent (w/v) of sucrose, wherein the pH of said aqueous composition is pH 8 plus or minus 0.5.
3. The method of claim 2 , wherein the aqueous composition comprises about 10 mg/ml of the polypeptide of SEQ ID NO:1, about 5% (w/v) of mannitol, about 0.5% (w/v) of sucrose, about 20 mM sodium citrate, about 5 mM Tris, about 0.1 mM zinc sulfate, about 0.01% (w/v) TWEEN 80, wherein the pH of said aqueous composition is pH 8 plus or minus 0.5.
4. The method of claim 3 , wherein the aqueous composition consists essentially of 10 mg/ml of the polypeptide of SEQ ID NO:1, 5% (w/v) of mannitol, 0.5% (w/v) of sucrose, 20mM sodium citrate, 5 mM Tris, 0.1 mM zinc sulfate, 0.01% (w/v) TWEEN 80, wherein the pH of said aqueous composition is pH 8 plus or minus 0.5.