Formulations of human tissue kallikrein-1 for parenteral delivery and related methods

Abstract

Provided are high concentration compositions of tissue kallikrein-1 (KLK1) and methods of parenterally administering such compositions to a subject in need thereof, where absorption into the circulation via, for example, intravenous or subcutaneous administration improves systemic pharmacokinetics, bioavailability, safety, and/or convenience relative to intravenous or other forms of administration. Also provided are recombinant human KLK1 (rhKLK1) polypeptides that can be readily concentrated to high protein concentrations, and substantially pure compositions thereof.

What is claimed is: 
     
         1 . A composition formulated for parenteral administration, the composition comprising a human tissue kallikrein-1 (hKLK1) polypeptide and a pharmaceutically acceptable carrier, wherein the concentration of the hKLK1 polypeptide in the composition is greater than about 5 mg/mL. 
     
     
         2 . The composition of  claim 1 , wherein the hKLK1 polypeptide has a pI of less than about 5 and a sialic acid content of at least about 4 moles per mole protein. 
     
     
         3 . The composition of  claim 1 , wherein the hKLK1 concentration is greater than about 10 mg/mL. 
     
     
         4 . The composition of  claim 1 , wherein the hKLK1 concentration is greater than about 25 mg/mL. 
     
     
         5 . The composition of  claim 1 , wherein the composition is substantially free of aggregates (greater than about 95% appearing as a single peak by SEC HPLC). 
     
     
         6 . The composition of  claim 1 , wherein the composition has endotoxin levels of less than about 1 EU/mg protein, host cell protein of less than about 100 ng/mg protein, and host cell DNA of less than about 10 pg/mg protein. 
     
     
         7 . The composition of  claim 1 , where the hKLK1 comprises an amino acid sequence with at least about 95% sequence identity to residues 25-262 of SEQ ID NO:1 or SEQ ID NO:2 and has serine protease activity. 
     
     
         8 . The composition of  claim 7 , where the serine protease activity is characterized by the ability to release kallidin from a higher molecular weight precursor. 
     
     
         9 . The composition of  claim 7 , where the hKLK1 polypeptide comprises E145 and/or A188, relative to SEQ ID NO:1. 
     
     
         10 . The composition of  claim 7 , where the hKLK1 polypeptide comprises Q145 and/or V188, relative to SEQ ID NO:1. 
     
     
         11 . The composition of  claim 1 , where the hKLK1 polypeptide comprises of residues 25-262 of SEQ ID NO:2. 
     
     
         12 . The composition of  claim 1 , where the hKLK1 polypeptide consists of residues 25-262 of SEQ ID NO:2. 
     
     
         13 . A method of treating a subject in need thereof, the method comprising parenterally administering to the subject a composition of  claim 1  and thereby treating the subject. 
     
     
         14 . The method of  claim 13 , where the composition is administered subcutaneously. 
     
     
         15 . The method of  claim 13 , where the composition is administered intravenously. 
     
     
         16 . The method of  claim 14  where administering the composition subcutaneously produces improved systemic pharmacokinetics relative to intravenously administration of the composition. 
     
     
         17 . The method of  claim 16 , where the improved pharmacokinetics comprises increased apparent half-life. 
     
     
         18 . The method of  claim 16 , wherein the improved pharmacokinetics comprises decreased Tmax. 
     
     
         19 . The method of  claim 16 , where the improved pharmacokinetics comprises increased Cmax. 
     
     
         20 . The method of  claim 16 , where the improved pharmacokinetics comprises increased absorption rate. 
     
     
         21 . A device comprising a composition of  claim 1 , wherein the device is suitable for parenteral delivery of the composition.