Antigen binding proteins to proprotein convertase subtilisin kexin type 9 (pcsk9)

Abstract

Antigen binding proteins that interact with Proprotein Convertase Subtilisin Kexin Type 9 (PCSK9) are described. Methods of treating hypercholesterolemia and other disorders by administering a pharmaceutically effective amount of an antigen binding protein to PCSK9 are described. Methods of detecting the amount of PCSK9 in a sample using an antigen binding protein to PCSK9 are described.

1 . An isolated neutralizing antigen binding protein that binds to a PCSK9 protein comprising the amino acid sequence of SEQ ID NO: 1, wherein the neutralizing antigen binding protein decreases the LDLR lowering effect of PCSK9 on LDLR. 
     
     
         2 . The isolated neutralizing antigen binding protein of  claim 1 , wherein the antigen binding protein is a LDLR non-competitive neutralizing antigen binding protein. 
     
     
         3 . The isolated neutralizing antigen binding protein of  claim 1 , wherein the antigen binding protein is a LDLR competitive neutralizing antigen binding protein. 
     
     
         4 . An antigen binding protein that selectively binds to PCSK9, wherein said antigen binding protein binds to PCSK9 with a K d  that is less than 100 pM. 
     
     
         5 . The antigen binding protein of  claim 4 , wherein the antigen binding protein binds with a K d  that is less than 10 pM. 
     
     
         6 . The antigen binding protein of  claim 4 , wherein the antigen binding protein binds with a K d  that is less than 5 pM. 
     
     
         7 . An isolated antigen binding protein that binds to a PCSK9 protein of SEQ ID NO: 1, wherein the binding between said isolated antigen binding protein and a variant PCSK9 protein is less than 50% of the binding between the isolated antigen binding protein and the PCSK9 protein of SEQ ID NO: 1. 
     
     
         8 . The isolated human antigen binding protein of  claim 7 , wherein the variant PCSK9 protein comprises at least one mutation of a residue at a position selected from the group consisting of 207, 208, 185, 181, 439, 513, 538, 539, 132, 351, 390, 413, 582, 162, 164, 167, 123, 129, 311, 313, 337, 519, 521, and 554, as shown in SEQ ID NO: 1. 
     
     
         9 . The isolated human antigen binding protein of  claim 8 , wherein the at least one mutation selected from the group consisting of R207E, D208R, E181R, R185E, R439E, E513R, V538R, E539R, T132R, S351R, A390R, A413R, and E582R 
     
     
         10 . The isolated human antigen binding protein of  claim 8 , wherein the at least one mutation is selected from the group consisting of D162R, R164E, E167R, S123R, E129R, A311R, D313R, D337R, R519E, H521R, and Q554R. 
     
     
         11 . An antigen binding protein that binds to a PCSK 9 protein of SEQ ID NO: 303 in a first manner, wherein the antigen binding protein binds to a variant of PCSK9 in a second manner, wherein said PCSK9 variant has at least one point mutation at a position selected from the group consisting of: 207, 208, 185, 181, 439, 513, 538, 539, 132, 351, 390, 413, 582, 162, 164, 167, 123, 129, 311, 313, 337, 519, 521, and 554 of SEQ ID NO: 303, wherein the first manner comprises a first EC50, a first Bmax, or a first EC50 and a first Bmax, wherein the second manner comprises a second EC50, a second Bmax, or a second EC50 and a second Bmax, and wherein a value for the first manner is different from a value for the second manner. 
     
     
         12 . The antigen binding protein of  claim 11 , wherein the first manner comprises a first EC50, wherein the second manner involves a second EC50, and wherein the point mutation is selected from the group consisting of: R207E, D208R, E181R, R185E, R439E, E513R, V538R, E539R, T132R, S351R, A390R, A413R, and E582R. 
     
     
         13 . The antigen binding protein of  claim 12 , wherein the first EC50 is at least 20% different from the second EC50. 
     
     
         14 . The antigen binding protein of  claim 12 , wherein the first EC50 is at least 50% different from the second EC50. 
     
     
         15 . The antigen binding protein of  claim 12 , wherein the second EC50 is a larger numerical value than the first EC50. 
     
     
         16 . The antigen binding protein of  claim 12 , wherein the first EC50 is determined by multiplex bead binding assay. 
     
     
         17 . The antigen binding protein of  claim 12 , wherein the second EC50 is greater than 1 uM. 
     
     
         18 . The antigen binding protein of  claim 11 , wherein the antigen binding protein is a neutralizing antigen binding protein. 
     
     
         19 . The antigen binding protein of  claim 18 , wherein the neutralizing antigen binding protein is a competitive neutralizing antigen binding protein. 
     
     
         20 . The antigen binding protein of  claim 18 , wherein the neutralizing antigen binding protein is a non-competitive neutralizing antigen binding protein. 
     
     
         21 . The antigen binding protein of  claim 11 , wherein the first manner comprises a first Bmax, wherein the second manner comprises a second Bmax that is different from the first Bmax, and wherein said PCSK9 variant has at least one point mutation selected from the group consisting of: D162R, R164E, E167R, S123R, E129R, A311R, D313R, D337R, R519E, H521R, and Q554R. 
     
     
         22 . The antigen binding protein of  claim 21 , wherein the second Bmax is about 10% of the first Bmax. 
     
     
         23 . The antigen binding protein of  claim 21 , wherein the first Bmax is at least 20% different from the second Bmax. 
     
     
         24 . The antigen binding protein of  claim 21 , wherein the first Bmax is at least 50% different from the second Bmax. 
     
     
         25 . An isolated antibody that binds to PCSK9 at a location that overlaps with a location that LDLR binds to PCSK9. 
     
     
         26 . A neutralizing antibody that binds to PCSK9 and reduces a low density lipoprotein receptor (LDLR) lowering effect of PCSK9 on LDLR. 
     
     
         27 . A neutralizing antibody that binds to PCSK9, wherein the antibody binds to PCSK9 at a location within residues 31-447 of SEQ ID NO: 3. 
     
     
         28 . The antibody of  claim 27 , wherein the antibody binds to an epitope within residues 31-447 of SEQ ID NO: 3. 
     
     
         29 . A pharmaceutical composition comprising at least one antigen binding protein according to  claim 1 , and a pharmaceutically acceptable excipient. 
     
     
         30 . A nucleic acid molecule encoding the antigen binding protein according to  claim 1 . 
     
     
         31 . A method of making an antigen binding protein that binds to a PCSK9 protein comprising the amino acid sequence of SEQ ID NO: 1, wherein the antigen binding protein decreases the LDLR lowering effect of PCSK9 on LDLR, said method comprising:
 providing a host cell comprising a nucleic acid sequence that encodes the antigen binding protein; and   maintaining the host cell under conditions in which the antigen binding protein is expressed.   
     
     
         32 . A method of lowering serum cholesterol level in a subject, said method comprising administering to a subject an effective amount of an isolated neutralizing antigen binding protein that binds to a PCSK9 protein comprising the amino acid sequence of SEQ ID NO: 1, wherein the neutralizing antigen binding protein decreases the LDLR lowering effect of PCSK9 on LDLR. 
     
     
         33 . A method for treating or preventing a condition associated with an elevated serum cholesterol level in a patient, comprising administering to a patient in need thereof an effective amount of an isolated neutralizing antigen binding protein that binds to a PCSK9 protein comprising the amino acid sequence of SEQ ID NO: 1, wherein the neutralizing antigen binding protein decreases the LDLR lowering effect of PCSK9 on LDLR. 
     
     
         34 . A method for treating or preventing a condition associated with elevated serum cholesterol levels in a subject, said method comprising administering to a subject in need thereof an effective amount of an isolated neutralizing antigen binding protein simultaneously or sequentially with an agent that elevates the availability of LDLR protein, wherein the isolated antigen binding protein binds to a PCSK9 protein comprising the amino acid sequence of SEQ ID NO: 1, wherein the neutralizing antigen binding protein decreases the LDLR lowering effect of PCSK9 on LDLR. 
     
     
         35 . The method of  claim 34 , wherein the agent that elevates the availability of LDLR protein comprises a statin. 
     
     
         36 . The method of  claim 35 , wherein the statin is selected from the group consisting of atorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin, pravastatin, rosuvastatin, simvastatin, and some combination thereof.