US2013323222A1PendingUtilityA1

Human tissue kallikrein 1 glycosylation isoforms

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Assignee: CHARLES MATTHEWPriority: Jun 4, 2012Filed: Jun 4, 2013Published: Dec 5, 2013
Est. expiryJun 4, 2032(~5.9 yrs left)· nominal 20-yr term from priority
Inventors:Matthew CharlesTadeusz KolodkaMark Williams
A61P 3/10C12N 9/96C12N 9/6445C12Y 304/21035A61P 3/00A61K 38/4853
51
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Claims

Abstract

Provided are preparations of tissue kallikrein-1 (KLK1) glycoforms having a defined number of oligosaccharide units per KLK1 molecule. Also provided are mixtures of such glycoforms, pharmaceutical compositions containing such glycoforms or mixtures thereof, methods of obtaining the rhKLK1 glycoforms, and therapeutic uses thereof

Claims

exact text as granted — not AI-modified
What is claimed is: 
     
         1 . A composition, comprising a first tissue kallikrein-1 (KLK1) polypeptide and a second tissue kallikrein-1 (KLK1) polypeptide:
 wherein the first KLK1 polypeptide has three glycans attached at three different positions per polypeptide and the second KLK1 polypeptide has two glycans attached at two different positions per polypeptide; and   wherein the first KLK1 polypeptide and the second KLK1 polypeptides are present in the composition in a ratio ranging from about 45:55 to about 55:45.   
     
     
         2 . The composition of  claim 1 , where one or more of said glycans are N-linked glycans. 
     
     
         3 . The composition of  claim 1 , where one or more of the glycans are attached at amino acid residues 78, 84, or 141 of KLK1 as defined by SEQ ID NO:l. 
     
     
         4 . The composition of  claim 3 , where the three glycans of the first KLK1 polypeptide are N-linked glycans at residues 78, 84, and 141. 
     
     
         5 . The composition of  claim 3 , where the two glycans of the second KLK1 polypeptide are N-linked glycans at residues 78 and 84 but not 141. 
     
     
         6 . The composition of  claim 1 , wherein the ratio of first KLK1 polypeptide and the second KLK1 polypeptide in the composition is about 50:50. 
     
     
         7 . A composition comprising a triple glycosylated isoform of a tissue kallikrein-1 (KLK1) polypeptide and a double glycosylated isoform of a tissue kallikrein-1 (KLK1) polypeptide, wherein the triple glycosylated isoform of the KLK1 polypeptide and the double glycosylated isoform of the KLK1 polypeptide are present in the composition in a ratio ranging from about 45:55 to about 55:45. 
     
     
         8 . The composition of  claim 7 , wherein the triple glycosylated isoform comprises N-linked glycans at amino acid residues 78, 84, and 141 of KLK1, as defined by SEQ ID NO:1. 
     
     
         9 . The composition of  claim 7 , wherein the double glycosylated isoform comprises N-linked glycans at amino acid residues 78 and 84, but not at amino acid residue 141 of KLK1, as defined by SEQ ID NO:1. 
     
     
         10 . The composition of  claim 7 , wherein the triple glycosylated isoform and the double glycosylated isoform of KLK1 are present in the composition in a ratio of about 50:50. 
     
     
         11 . The composition of  claim 1 , wherein said KLK1 polypeptide(s) comprise a human tissue kallikrein-1 (hKLK1) polypeptide. 
     
     
         12 . The composition of  claim 1 , wherein said KLK1 polypeptide(s) comprise amino acid residues 78-141 of SEQ ID NO:1 or amino acids residues 78-141 SEQ ID NO:2. 
     
     
         13 . The composition of  claim 1 , wherein said KLK1 polypeptide(s) comprise amino acid residues 25-262 of SEQ ID NO:1 or amino acid residues 25-262 of SEQ ID NO:2 
     
     
         14 . The composition of  claim 1 , wherein said KLK1 polypeptide(s) comprise an amino acid sequence having at least about 95% sequence identity to SEQ ID NO:1 or SEQ ID NO:2. 
     
     
         15 . The composition of  claim 1 , wherein said KLK1 polypeptide(s) comprise an amino acid sequence having at least about 95% sequence identity to amino acid residues 25-262 of SEQ ID NO:1 or SEQ ID NO:2. 
     
     
         16 . The composition of  claim 1 , wherein said KLK1 polypeptide(s) comprise an amino acid sequence having at least about 95% sequence identity to amino acid residues 25-262 of SEQ ID NO:2, and wherein said KLK1 polypeptide(s) comprises E145 and/or A188. 
     
     
         17 . The composition of  claim 1 , wherein said KLK1 polypeptide(s) comprise an amino acid sequence having at least about 95% sequence identity to amino acid residues 25-262 of SEQ ID NO:2, and wherein said KLK1 polypeptide(s) comprises Q145 and/or V188. 
     
     
         18 . The composition of  claim 1 , wherein the KLK1 polypeptide(s) comprises SEQ ID NO:1 or SEQ ID NO:2. 
     
     
         19 . The composition of  claim 1 , wherein the KLK1 polypeptide(s) comprises residues 25-262 of SEQ ID NO:1 or SEQ ID NO:2. 
     
     
         20 . The composition of  claim 1 , wherein the KLK1 polypeptide(s) consists of amino acids residues 25-262 of SEQ ID NO:1 or amino acid residues 25-262 of SEQ ID NO:2. 
     
     
         21 . The composition of  claim 1 , further comprising a pharmaceutically acceptable diluent, adjuvant, or carrier. 
     
     
         22 . The composition of  claim 1 , where the composition is substantially free of other glycosylated isoforms (glycoforms) of KLK1. 
     
     
         23 . The composition of  claim 1 , where the composition has endotoxin levels of less than about 1 EU/mg protein, host cell protein of less than about 100 ng/mg total protein, host cell DNA of less than about 10 pg/mg total protein, and/or is substantially free of aggregates (greater than about 95% appearing as a single peak by SEC HPLC). 
     
     
         24 . A device comprising a composition of  claim 1 , wherein the device is suitable for delivering the composition subcutaneously. 
     
     
         25 . A method of treating a subject in need thereof, comprising administering to the subject a composition of  claim 1 .

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