COLLAGENASE FORMULATIONS AND METHODS OF PRODUCING THE SAME
Disclosed herein are improved collagenase-containing formulations and methods of preparing the same. The collagenase-containing formulations comprise a collagenase, about 30 mM to about 240 mM of a disaccharide, about 50 mM to about 800 mM of mannitol, and about 6 mM to about 10 mM of a Tris-HCl. Lyophilized and reconstituted formulations are also provided.
1 . A lyophilized formulation comprising:
a collagenase;
a disaccharide;
mannitol; and
Tris-HCl.
2 . The lyophilized formulation of claim 1 , wherein the collagenase is produced by recombinant techniques.
3 . The lyophilized formulation of claim 1 , wherein the disaccharide is sucrose or trehalose.
4 . The lyophilized formulation of claim 1 , wherein, prior to lyophilization, the formulation comprises:
a collagenase;
60 mM of sucrose;
225 mM of mannitol; and
10 mM of Tris-HCl, and
the formulation has a pH of about 8.5.
5 . The lyophilized formulation of claim 1 , wherein the lyophilized formulation is stable at pressures above 380 pbar during lyophilization.
6 . The lyophilized formulation of claim 5 , wherein the lyophilized formulation is stable at a pressure of about 4000 pbar during lyophilization.
7 . The lyophilized formulation of claim 1 , wherein the lyophilized formulation is stable at:
(a) 2-8° C. for at least 36 months;
(b) 25° C. and 60% relative humidity for at least 36 months;
(c) 40° C. and 75% relative humidity for at least 6 months; or
(d) any combination of (a) to (c).
8 . The lyophilized formulation of claim 1 , wherein the lyophilized formulation is formed by a method comprising:
freezing the formulation at a temperature of about −25° C. to about −55° C. to form a frozen formulation; and
drying the frozen formulation at a temperature of about 25° C. to about 50° C. to form the lyophilized formulation.
9 . The lyophilized formulation of claim 8 , wherein the lyophilized formulation is formed by a method comprising:
freezing the formulation at a single temperature of about −25° C. to about −55° C. to form a frozen formulation; and
drying the frozen formulation at a single temperature of about 25° C. to about 50° C. to form the lyophilized formulation.
10 . The lyophilized formulation of claim 1 , wherein the lyophilized formulation is formed by a lyophilization method that is performed for less than 72 hours.
11 . The lyophilized formulation of claim 1 , wherein the lyophilized formulation is formed by a lyophilization method that is performed at a pressure of about 380 pbar to about 4000 pbar.
12 . The lyophilized formulation of claim 1 , in a unit-dose vial, multi-dose vial, cartridge, or syringe.
13 . A reconstituted formulation comprising:
a collagenase;
a disaccharide;
mannitol;
Tris-HCl;
calcium chloride; and
sodium chloride.
14 . The reconstituted formulation of claim 13 , wherein the collagenase is produced by recombinant techniques.
15 . The reconstituted formulation of claim 13 , wherein the disaccharide is sucrose or trehalose.
16 . The reconstituted formulation of claim 13 , wherein the reconstituted formulation is isotonic to human blood.
17 . A kit comprising:
a container comprising the lyophilized formulation of claim 1 ; and
a container comprising a sterile diluent comprising calcium chloride and sodium chloride.