Antigen binding proteins to proprotein convertase subtilisin kexin type 9 (PCSK9)

• US 9,493,576 B2
• Filed: 08/14/2014
• Issued: 11/15/2016
• Est. Priority Date: 08/23/2007
• Status: Active Grant

• Current Assignee: Amgen Inc.
• Original Assignee: Amgen Inc.
• Inventors: Jackson, Simon Mark, Walker, Nigel Pelham Clinton, Piper, Derek Evan, Shan, Bei, Shen, Wenyan, King, Chadwick Terence, Ketchem, Randal Robert, Mehlin, Christopher, Carabeo, Teresa Arazas
• Primary Examiner(s): WEN, SHARON X
• Application Number:
  US14/459,787
• Publication Number:
  US 20140357851A1
• Time in Patent Office: 824 Days
• US Class Current: 1/1
• 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. A monoclonal antibody that recognizes a conformational epitope on human PCSK9 comprising amino acid residues:

S153, R194, D238, D374, T377, and F379 of SEQ ID NO;

3, wherein the monoclonal antibody reduces binding between PCSK9 and EGFa domain of LDLR.

2. The monoclonal antibody of claim 1, wherein the monoclonal antibody is a neutralizing antibody.

3. The monoclonal antibody of claim 1 wherein the monoclonal antibody is a human antibody.

4. The monoclonal antibody of claim 1, wherein the monoclonal antibody is a humanized antibody.

5. The monoclonal antibody of claim 1, wherein the monoclonal antibody was produced by a mammalian cell.

6. The monoclonal antibody of claim 1, wherein the monoclonal antibody was produced by a CHO cell.

7. The monoclonal antibody of claim 1, wherein the monoclonal antibody binds to PCSK9 with a K_D of less than or equal to 5×

10<sup>−

9</sup> M.

8. The monoclonal antibody of claim 1, wherein the monoclonal antibody comprises a light chain region that comprises an amino acid sequence of SEQ ID NO:

157.

9. The monoclonal antibody of claim 1, wherein the monoclonal antibody comprises a heavy chain region that comprises an amino acid sequence of SEQ ID NO:

154.

10. The monoclonal antibody of claim 9, wherein the monoclonal antibody comprises a light chain region that comprises an amino acid sequence of SEQ ID NO:

157.

11. The monoclonal antibody of claim 10, further comprising a HCDR1 that is a HCDR1 in SEQ ID NO:

60 according to the definition of Chothia.

12. The monoclonal antibody of claim 1, further comprising a HCDR1 that is a HCDR1 in SEQ ID NO:

60 according to the definition of Chothia.

43. A pharmaceutical composition comprising:

a pharmaceutically acceptable carrier; and

the monoclonal antibody of any one of claims 1-42.

13. A monoclonal antibody that binds to amino acids in human PCSK9 of SEQ ID NO:

1, wherein the amino acids comprise S153, R194, D238, D374, T377, and F379 of SEQ ID NO;

3, and wherein the monoclonal antibody reduces binding between PCSK9 and EGFa domain of LDLR.

14. The monoclonal antibody of claim 13, wherein the monoclonal antibody is a neutralizing antibody.

15. The monoclonal antibody of claim 13, wherein the monoclonal antibody is a humanized antibody.

16. The monoclonal antibody of claim 13, wherein the monoclonal antibody was produced by a mammalian cell.

17. The monoclonal antibody of claim 13, wherein the monoclonal antibody was produced by a CHO cell.

18. The monoclonal antibody of claim 13, wherein the monoclonal antibody comprises a light chain region that comprises an amino acid sequence of SEQ ID NO:

157.

19. The monoclonal antibody of claim 13, wherein the monoclonal antibody comprises a heavy chain region that comprises an amino acid sequence of SEQ ID NO:

154.

20. The monoclonal antibody of claim 19, wherein the monoclonal antibody comprises a light chain region that comprises an amino acid sequence of SEQ ID NO:

157.

21. The monoclonal antibody of claim 20, further comprising a HCDR1 that is a HCDR1 in SEQ ID NO:

60 according to the definition of Chothia.

22. The monoclonal antibody of claim 13, further comprising a HCDR1 that is a HCDR1 in SEQ ID NO:

60 according to the definition of Chothia.

23. A monoclonal antibody that recognizes a conformational epitope on human PCSK9, wherein the conformational epitope comprises at least ten of the following residues:

S153, 1154, P155, R194, D238, A239, 1369, S372, D374, C375, T377, C378, F379, V380, or S381 of SEQ ID NO;

3, wherein the monoclonal antibody reduces binding between PCSK9 and EGFa domain of LDLR.

24. The monoclonal antibody of claim 23, wherein the conformational epitope comprises at least thirteen of the following residues:

S153, I154, P155, R194, D238, A239, I369, S372, D374, C375, T377, C378, F379, V380, or S381 of SEQ ID NO;

3.

25. The monoclonal antibody of claim 23, wherein the monoclonal antibody binds to at least S381.

26. The monoclonal antibody of claim 23, wherein the monoclonal antibody is a humanized antibody.

27. The monoclonal antibody of claim 23, wherein the monoclonal antibody was produced by a mammalian cell.

28. The monoclonal antibody of claim 23, wherein the monoclonal antibody was produced by a CHO cell.

29. The monoclonal antibody of claim 23, wherein the monoclonal antibody is a neutralizing antibody.

30. The monoclonal antibody of claim 23, wherein the monoclonal antibody comprises a light chain region that comprises an amino acid sequence of SEQ ID NO:

157.

31. The monoclonal antibody of claim 23, wherein the monoclonal antibody comprises a heavy chain region that comprises an amino acid sequence of SEQ ID NO:

154.

32. The monoclonal antibody of claim 31, wherein the monoclonal antibody comprises a light chain region that comprises an amino acid sequence of SEQ ID NO:

157.

33. A monoclonal antibody that binds to amino acids in human PCSK9, wherein the amino acids comprise ten or more of the following amino acids:

S153, I154, P155, R194, D238, A239, I369, S372, D374, C375, T377, C378, F379, V380, or S381 of SEQ ID NO;

3, and wherein the monoclonal antibody reduces binding between PCSK9 and EGFa domain of LDLR.

34. The monoclonal antibody of claim 33, wherein the monoclonal antibody binds thirteen or more of the following amino acids:

S153, I154, P155, R194, D238, A239, I369, S372, D374, C375, T377, C378, F379, V380, or S381 of SEQ ID NO;

3.

35. The monoclonal antibody of claim 33, wherein the monoclonal antibody binds to at least S381.

36. The monoclonal antibody of claim 33 wherein the monoclonal antibody is a humanized antibody.

37. The monoclonal antibody of claim 33, wherein the monoclonal antibody was produced by a mammalian cell.

38. The monoclonal antibody of claim 33, wherein the monoclonal antibody was produced by a CHO cell.

39. The monoclonal antibody of claim 33, wherein the monoclonal antibody is a neutralizing antibody.

40. The monoclonal antibody of claim 33, wherein the monoclonal antibody comprises a light chain region that comprises an amino acid sequence of SEQ ID NO:

157.

41. The monoclonal antibody of claim 33, wherein the monoclonal antibody comprises a heavy chain region that comprises an amino acid sequence of SEQ ID NO:

154.

42. The monoclonal antibody of claim 41, wherein the monoclonal antibody comprises a light chain region that comprises an amino acid sequence of SEQ ID NO:

157.