ANTIGEN BINDING PROTEINS TO PROPROTEIN CONVERTASE SUBTILISIN KEXIN TYPE 9 (PCSK9)

• US 20110027287A1
• Filed: 10/12/2010
• Published: 02/03/2011
• Est. Priority Date: 08/23/2007
• Status: Abandoned Application

• Current Assignee: Amgen Inc.
• Original Assignee: Amgen Inc.
• Inventors: Jackson, Simon Mark, Walker, Nigel Pelham Clinton, Piper, Derek Evan, Shan, Bei, Shen, Wenyan, Chan, Joyce Chi Yee, King, Chadwick Terence, Ketchem, Randal Robert, Mehlin, Christopher, Carabeo, Teresa Arazas, Cao, Qiong
• Application Number:
  US12/903,084
• Publication Number:
  US 20110027287A1
• Time in Patent Office: Days
• US Class Current: 424/139.1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/22 of acyclic acids, e.g. pravastatin
• CPC Class Codes: A61K 31/366 having six-membered rings, e.g. delta-lactones
• CPC Class Codes: A61K 31/40 having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
• CPC Class Codes: A61K 31/405 Indole-alkanecarboxylic acids; Derivatives thereof, e.g. tryptophan, indomethacin
• CPC Class Codes: A61K 31/44 Non condensed pyridines; Hydrogenated derivatives thereof
• CPC Class Codes: A61K 31/47 Quinolines; Isoquinolines
• CPC Class Codes: A61K 31/505 Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
• CPC Class Codes: A61K 31/66 Phosphorus compounds
• CPC Class Codes: A61K 39/395 Antibodies agglutinins A61K38/36 ; as drug carriers A61K47/50; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07K 16/40 : against enzymes
• CPC Class Codes: C07K 2299/00 : Coordinates from 3D structures of peptides, e.g. proteins or enzymes
• CPC Class Codes: C07K 2317/14 : Specific host cells or culture conditions, e.g. components, pH or temperature
• CPC Class Codes: C07K 2317/34 : Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/76 : Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 : Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C12N 15/1137 : against enzymes viral enzymes C12N15/1131; receptors C12N15/1138

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.

Claims

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.

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.

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.

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.