Lyophilization of synthetic liposomal pulmonary surfactant

1. A process of making a lyophilized synthetic pulmonary surfactant having a reduced or eliminated cake levitation during the process, the process comprising:

providing to a lyophilizing chamber a pre-lyophilization mixture comprising dipalmitoyl phosphatidylcholine (DPPC), palmitoyloleoyl phosphatidylglycerol (POPG), palmitic acid and a synthetic peptide with SEQ ID NO:1 (KL4 polypeptide) dispersed in a solvent having an organic solvent in a range of between 3% (v/v) and 20% (v/v) of the total volume of the pre-lyophilization mixture with a remainder being water and/or buffer, wherein the pre-lyophilization mixture is filled in a container;

lowering a temperature inside the lyophilizing chamber to begin chilling and solidifying the pre-lyophilization mixture in a freezing phase; and

conducting an annealing phase prior to a primary drying phase and thereby reducing or eliminating cake levitation in the lyophilized synthetic pulmonary surfactant wherein the lyophilized synthetic pulmonary surfactant has a specific surface area of at least 2.7 m 2 /g.

2. The process of claim 1 , the process comprising:

conducting the freezing phase in a process of lowering the temperature inside the lyophilizing chamber, wherein the pre-lyophilization mixture is chilled to a first temperature below −45° C. at a rate between 0.1 and 1.0° C./min and holding the pre-lyophilization mixture at the first temperature for a first period of time sufficient to solidify at least 76% of the solvent to form a first solidified mixture;

conducting the annealing phase and thereby reducing or eliminating cake levitation of the first solidified mixture, wherein the first solidified mixture is (i) heated to a second temperature selected to reduce or eliminate levitation of the first solidified mixture, (ii) held at the second temperature for a second period of time sufficient to reduce or eliminate levitation of the first solidified mixture, and (iii) chilled to a third temperature below −45° C. at a rate between 0.1 to 1.0° C./min to form a second solidified mixture, wherein the second solidified mixture is held at the third temperature for a third period of time sufficient to promote separation of unfrozen organic solvent from the second solidified mixture and thereby achieve a migration of the unfrozen organic solvent to an interface between the container and the second solidified mixture;

conducting a primary drying phase at a reduced pressure of 30 mT or higher, wherein the second solidified mixture is held at the fourth temperature for a fourth period of time sufficient to sublimate at least 10% of the organic solvent, followed by heating to a fourth temperature sufficient to keep the second solidified mixture from levitating in the container and retaining a structure established during the annealing phase, and further held at the fourth temperature for a fifth period of time sufficient to sublimate at least 85% of the solvent and thereby forming a third solidified mixture; and

conducting a secondary drying phase at the reduced pressure for a sixth period of time sufficient to produce the lyophilized synthetic pulmonary surfactant having a residual solvent content of at most 2%.

3. The process of claim 2 , wherein a ratio of the pre-lyophilization mixture's volume in the container to the container's volume is from about 28% to about 68%.

4. The process of claim 2 , the process comprising:

providing the pre-lyophilization mixture wherein the organic solvent in the range from about 3% to about 15%.

5. The process of claim 2 , the process comprising:

providing the pre-lyophilization mixture wherein the organic solvent in the range from about 5% to about 10%.

6. The process of claim 2 , the process comprising:

providing the pre-lyophilization mixture wherein the organic solvent in the range from about 7% to about 10%.

7. The process of claim 2 , the process comprising:

conducting the freezing phase, wherein the pre-lyophilization mixture is chilled to the first temperature −50° C.±3° C. at the rate between 0.1 and 1.0° C./min;

conducting the annealing phase, wherein the first solidified mixture is (i) heated to the second temperature of −22° C.±3° C. at a rate of 0.1 to 1.0° C./min, (ii) held at the second temperature for the second period of time between 4 and 8 hours, (iii) chilled to the third temperature of −50° C.±3° C. at a rate between 0.1 to 1.0° C./min; and (iv) held at the third temperature for the third period of time for about 3 to 8 hours;

conducting the primary drying phase at a pressure of 40 mT±10 mT and temperature of −25° C.±3° C. ramped up from −50° C.±3° C., and further held at −25° C.±13° C. for 60 to 140 hours; and

conducting the secondary drying phase at a pressure of 40 mT±10 mT and temperature of 25° C.±3° C. at 0.5° C./min ramped up from −25° C.±3° C. and held at 25° C.±3° C. for 4 to 10 hours.

8. The process of claim 2 , wherein the specific surface area is in the range from about 3.7 m 2 /g to about 2.7 m 2 /g.

9. The process of claim 2 , wherein the lyophilized synthetic pulmonary surfactant has porosity above 40% by volume of a total area of the lyophilized synthetic pulmonary surfactant.

10. A lyophilized synthetic pulmonary surfactant composition comprising a synthetic peptide with SEQ ID NO:1 (KL4 polypeptide), dipalmitoyl phosphatidylcholine (DPPC), palmitoyloleoyl phosphatidylglycerol (POPG) and palmitic acid, wherein the lyophilized synthetic pulmonary surfactant composition has a specific surface area of at least 2.7 m 2 /g.

11. The lyophilized synthetic pulmonary surfactant of claim 10 , wherein the specific surface area is in the range from about 3.7 m 2 /g to about 2.7 m 2 /g.

12. The lyophilized synthetic pulmonary surfactant of claim 10 , wherein the lyophilized synthetic pulmonary surfactant has porosity above 40% by volume of a total area of the lyophilized synthetic pulmonary surfactant.