Methods To Produce Lung Surfactant Formulations Via Lyophilization And Formulations And Uses Thereof

Abstract

Lyophilized lung surfactant compositions having reduced viscosity upon reconstitution are described. These compositions are produced by a method including solvent dissolution and lyophilization.

1 . A lyophilized lung surfactant composition comprising:
 one or more phospholipids, at least one or more of a fatty acid, a fatty alcohol or a fatty acid ester, and a lung surfactant polypeptide having at least 10 amino acid residues and represented by a formula:
   (Z a U b ) c Z d    
   
       wherein Z represents a hydrophilic amino acid residue and U represents a hydrophobic amino acid residue; wherein each Z is independently R, D, E or K; and each U is independently V, I, L, C, Y, or F; and wherein a is an integer with an average value of about 1 to about 5; b is an integer with an average value of about 3 to about 20; c is an integer of about 1 to about 10; and d is an integer of about 1 to about 3, wherein the lyophilized lung surfactant composition has a reduced viscosity upon reconstitution as compared to a viscosity of an identical liquid lung surfactant which was not lyophilized, such that the reduced viscosity is equal to an apparent viscosity of less than about 50 cP at a concentration of 30 mg total phospholipid (TPL)/ml in Tris-Ac buffer at pH 7.6 containing 130 mM NaCl when measured at a temperature of 25° C. and a shear rate of 157 sec −1  using a rheometer fitted with a 40 mm/1° acrylic cone, and a step flow procedure with a linear increase in shear rate with a time of 0 to 200 sec −1  over approximately 3 minutes followed by a linear decrease in shear rate with a time of 200 sec −1  to 0 sec −1  over approximately 3 minutes, and wherein the lyophilized lung surfactant composition is produced by a method, the method comprising:
 combining the lung surfactant polypeptide, one or more phospholipids, and at least one or more of a fatty acid, a fatty alcohol or a fatty acid ester in an organic solvent system, said system comprising an organic solvent in an amount sufficient to (i) solubilize the lung surfactant polypeptide and lipids to form a substantially homogeneous liquid mixture and (ii) reduce viscosity of the lyophilized lung surfactant upon reconstitution, wherein the organic solvent system comprises from about 30% by volume to about 70% by volume of the organic solvent with the remainder being aqueous solvent and the organic solvent comprises a lower oxyhydrocarbon; and 
 lyophilizing the substantially homogeneous liquid mixture to obtain the lyophilized lung surfactant composition. 
 
     
     
         2 . The lyophilized lung surfactant composition of  claim 1  that comprises SEQ. ID. NO: 1 (KL4). 
     
     
         3 . The lyophilized lung surfactant composition of  claim 1 , wherein the reduced viscosity equals to an apparent viscosity of less than about 30 cP at a concentration of 30 mg total phospholipid/ml in Tris-Ac buffer at pH 7.6 containing 130 mM NaCl when measured at a temperature of 25° C. and a shear rate of 157 sec −1  using a rheometer fitted with a 40 mm/1° acrylic cone, and a step flow procedure with a linear increase in shear rate with a time of 0 to 200 sec −1  over approximately 3 minutes followed by a linear decrease in shear rate with a time of 200 sec −1  to 0 sec −1  over approximately 3 minutes. 
     
     
         4 . The lyophilized lung surfactant composition of  claim 3  that comprises SEQ. ID. NO:1 (KL4). 
     
     
         5 . The lyophilized lung surfactant composition of  claim 3  that comprises SEQ ID NO: 1 (KL4 polypeptide), dipalmitoyl phosphatidylcholine (DPPC), palmitoyloleoyl phosphatidylglycerol (POPG) and palmitic acid. 
     
     
         6 . The lyophilized lung surfactant composition of  claim 1 , wherein the reduced viscosity equals to an apparent viscosity of less than about 15 cP at a concentration of 30 mg total phospholipid/ml in Tris-Ac buffer at pH 7.6 containing 130 mM NaCl when measured at a temperature of 25° C. and a shear rate of 157 sec −1  using a rheometer fitted with a 40 mm/1° acrylic cone, and a step flow procedure with a linear increase in shear rate with a time of 0 to 200 sec −1  over approximately 3 minutes followed by a linear decrease in shear rate with a time of 200 sec −1  to 0 sec −1  over approximately 3 minutes. 
     
     
         7 . The lyophilized lung surfactant composition of  claim 6  that comprises SEQ. ID. NO: 1 (KL4). 
     
     
         8 . The lyophilized lung surfactant composition of  claim 6  that comprises SEQ ID NO: 1 (KL4 polypeptide), dipalmitoyl phosphatidylcholine (DPPC), palmitoyloleoyl phosphatidylglycerol (POPG) and palmitic acid. 
     
     
         9 . The lyophilized lung surfactant composition of  claim 1 , wherein the reduced viscosity equals to an apparent viscosity of less than about 30 cP at a concentration of 30 mg total phospholipid/ml in Tris-Ac buffer at pH 7.6 containing 130 mM NaCl when measured at a temperature of 25° C. and a shear rate of 157 sec −1  using a rheometer fitted with a 40 mm/1° acrylic cone, and a step flow procedure with a linear increase in shear rate with a time of 0 to 200 sec −1  over approximately 3 minutes followed by a linear decrease in shear rate with a time of 200 sec −1  to 0 sec −1  over approximately 3 minutes. 
     
     
         10 . The lyophilized lung surfactant composition of  claim 9  that comprises SEQ. ID. NO: 1 (KL4). 
     
     
         11 . The lyophilized lung surfactant composition of  claim 9  that comprises SEQ ID NO: 1 (KL4 polypeptide), dipalmitoyl phosphatidylcholine (DPPC), palmitoyloleoyl phosphatidylglycerol (POPG) and palmitic acid.