Nucleic acid molecules encoding a humanized antibody

• US 8,796,439 B2
• Filed: 08/07/2012
• Issued: 08/05/2014
• Est. Priority Date: 07/14/2006
• Status: Active Grant

• Current Assignee: AC Immune SA, Genetec Incorporated (Noritsu Koki Co., Ltd.)
• Original Assignee: AC Immune SA, Genetec Incorporated (Noritsu Koki Co., Ltd.)
• Inventors: Pfeifer, Andrea, Pihlgren, Maria, Muhs, Andreas, Watts, Ryan
• Primary Examiner(s): BALLARD, KIMBERLY
• Application Number:
  US13/568,995
• Publication Number:
  US 20120329149A1
• Time in Patent Office: 728 Days
• Field of Search: None
• US Class Current: 536/23.53
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• 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: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• 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 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: G01N 33/6896 Neurological disorders, e.g. Alzheimer's disease

Abstract

The present invention is related to chimeric and humanized antibody and to methods and compositions for the therapeutic and diagnostic use in the treatment of amyloidosis, a group of disorders and abnormalities associated with amyloid protein such as Alzheimer'"'"'s disease.

Claims

1. A nucleic acid molecule comprising a nucleotide sequence encoding a Light Chain Variable Region (LCVR) or a fragment thereof of an antibody which is capable of specifically binding to beta-amyloid 1-40 and to monomeric, oligomeric or polymeric forms of beta-amyloid 1-42, wherein said LCVR or fragment thereof comprises:

(i) human- or primate-derived framework regions,

(ii) an LCVR CDR1 comprising the amino acid sequence of SEQ ID NO;

4,

(iii) an LCVR CDR2 comprising the amino acid sequence of SEQ ID NO;

5 or the amino acid sequence of SEQ ID NO;

5 with one amino acid substitution, and

(iv) an LCVR CDR3 comprising the amino acid sequence of SEQ ID NO;

6,

and wherein said LCVR or fragment thereof comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO;

12.

4. The nucleic acid molecule of claim 1 comprising the nucleotide sequence of SEQ ID NO:

21 encoding a light chain variable region.

5. The nucleic acid molecule of claim 4 further comprising the nucleotide sequence of SEQ ID NO:

23 encoding a light chain constant region.

6. The nucleic acid molecule of claim 1, wherein the LCVR CDR2 comprises the amino acid sequence RVSNRFS (SEQ ID NO:

40) or KVSSRFS (SEQ ID NO;

41).

7. The nucleic acid molecule of claim 1 comprising the nucleotide sequence of SEQ ID NO:

22.

13. An expression vector comprising the nucleic acid molecule of any one of claims 1 to 3, 4 and 8.

14. A cell comprising the expression vector of claim 13.

15. An expression vector comprising the nucleic acid molecule of any one of claims 6-7.

16. A cell comprising the nucleic acid molecule of any one of claims 6-7.

17. The nucleic acid molecule of claim 1 which comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:

13.

20. An expression vector comprising the nucleic acid molecule of claim 17, 18, or 19.

21. A cell comprising the expression vector of claim 20.

22. An expression vector comprising the nucleic acid molecule of claim 1.

23. A cell comprising the expression vector of claim 22.

24. The cell of claim 23 further comprising an expression vector comprising a nucleic acid sequence encoding an amino acid sequence that is at least 98% identical to SEQ ID NO:

15.

27. A population of nucleic acid molecules comprising a first nucleic acid molecule and a second nucleic acid molecule, wherein the first nucleic acid molecule is the nucleic acid molecule of claim 1, and wherein the second nucleic acid molecule comprises a nucleotide sequence encoding an Heavy Chain Variable Region (HCVR) or a fragment thereof of an antibody which is capable of specifically binding to beta-amyloid 1-40 and to monomeric, oligomeric or polymeric forms of beta-amyloid 1-42, wherein said HCVR or fragment thereof comprises:

(i) human- or primate-derived framework regions,

(ii) an HCVR CDR1 comprising the amino acid sequence of SEQ ID NO;

1,

(iii) an HCVR CDR2 comprising the amino acid sequence of SEQ ID NO;

2, and

(iv) an HCVR CDR3 comprising the amino acid sequence of SEQ ID NO;

3.

29. A cell comprising the population of nucleic acid molecules of claim 27 or 28.

30. The population of nucleic acid molecules of claim 27, wherein the first nucleic acid molecule comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:

13 and the second nucleic acid molecule comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO;

16 with a C-terminal Lys deletion.

31. A cell comprising the population of nucleic acid molecules of claim 30.

32. A method of producing an antibody or a functional fragment thereof, which antibody specifically binds to beta-amyloid 1-40, and to monomeric, oligomeric or polymeric forms of beta-amyloid 1-42, wherein said method comprises expressing in a cell the nucleic acid molecule of claim 1.

2. A nucleic acid molecule comprising a nucleotide sequence encoding a Heavy Chain Variable Region (HCVR) or a fragment thereof of an antibody which is capable of specifically binding to beta-amyloid 1-40 and to monomeric, oligomeric or polymeric forms of beta-amyloid 1-42, wherein said HCVR or fragment thereof comprises:

(i) human- or primate-derived framework regions,

(ii) an HCVR CDR1 comprising the amino acid sequence of SEQ ID NO;

1,

(iii) an HCVR CDR2 comprising the amino acid sequence of SEQ ID NO;

2, and

(iv) an HCVR CDR3 comprising the amino acid sequence of SEQ ID NO;

3,

and wherein said HCVR or fragment thereof comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO;

15.

8. The nucleic acid molecule of claim 2 comprising the nucleotide sequence of SEQ ID NO:

24 encoding a heavy chain variable region.

9. The nucleic acid molecule of claim 8 further comprising the nucleotide sequence of SEQ ID NO:

26 encoding a heavy chain constant region.

10. The nucleic acid molecule of claim 2, which further comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:

17.

11. The nucleic acid molecule of claim 2 comprising the nucleotide sequence of SEQ ID NO:

25.

18. The nucleic acid molecule of claim 2 which comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:

16 with a C-terminal Lys deletion.

25. An expression vector comprising the nucleic acid molecule of claim 2.

26. A cell comprising the expression vector of claim 25.

28. A population of nucleic acid molecules comprising a first nucleic acid molecule and a second nucleic acid molecule, wherein the first nucleic acid molecule is the nucleic acid molecule of claim 2, and wherein the second nucleic acid molecule comprises a nucleotide sequence encoding a Light Chain Variable Region (LCVR) or a fragment thereof of an antibody which is capable of specifically binding to beta-amyloid 1-40 and to monomeric, oligomeric or polymeric forms of beta-amyloid 1-42, wherein said LCVR or fragment thereof comprises:

(i) human- or primate-derived framework regions,

(ii) an LCVR CDR1 comprising the amino acid sequence of SEQ ID NO;

4,

(iii) an LCVR CDR2 comprising the amino acid sequence of SEQ ID NO;

5 or the amino acid sequence of SEQ ID NO;

5 with one amino acid substitution, and

(vii) an LCVR CDR3 comprising the amino acid sequence of SEQ ID NO;

6.

33. A method of producing an antibody or a functional fragment thereof, which antibody specifically binds to beta-amyloid 1-40, and to monomeric, oligomeric or polymeric forms of beta-amyloid 1-42, wherein said method comprises expressing in a cell the nucleic acid molecule of claim 2.

3. A nucleic acid molecule comprising a nucleotide sequence encoding an antibody or a fragment thereof which antibody or fragment thereof is capable of specifically binding to beta-amyloid peptide 1-40 and to monomeric, oligomeric or polymeric forms of beta-amyloid 1-42, wherein said antibody or fragment thereof comprises:

(i) a Light Chain Variable Region (LCVR) comprising an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO;

12 and an Heavy Chain Variable Region (HCVR) comprising an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO;

15,

(ii) an HCVR CDR1 comprising the amino acid sequence of SEQ ID NO;

1,

(iii) an HCVR CDR2 comprising the amino acid sequence of SEQ ID NO;

2,

(iv) an HCVR CDR3 comprising the amino acid sequence of SEQ ID NO;

3,

(v) an LCVR CDR1 comprising the amino acid sequence of SEQ ID NO;

4,

(vi) an LCVR CDR2 comprising the amino acid sequence of SEQ ID NO;

5 or the amino acid sequence of SEQ ID NO;

5 with one amino acid substitution, and

(vii) an LCVR CDR3 comprising the amino acid sequence of SEQ ID NO;

6.

12. The nucleic acid molecule of claim 3 comprising a nucleotide sequence that hybridizes under stringent conditions to a complementary strand of any one of SEQ ID NOs:

21, 22, 24, or 25, wherein the stringent hybridization conditions comprise hybridization in 5×

SSPE, 1% SDS, 1×

Denhardts solution at 65°

C. and washing in 2×

SSC, 1% SDS and subsequently with 0.2×

SSC at 65°

C.

19. The nucleic acid molecule of claim 3, wherein the antibody or fragment thereof comprises a light chain comprising the amino acid sequence of SEQ ID NO:

13 and a heavy chain comprising the amino acid sequence of SEQ ID NO;

16 with a C-terminal Lys deletion.

34. A nucleic acid molecule comprising a nucleotide sequence encoding a Light Chain Variable Region (LCVR) or a fragment thereof of an antibody which is capable of specifically binding to beta-amyloid 1-40 and to monomeric, oligomeric or polymeric forms of beta-amyloid 1-42, wherein said LCVR comprises the amino acid sequence of SEQ ID NO:

12.

35. An expression vector comprising the nucleotide sequence of claim 34.

36. A cell comprising the expression vector of claim 35.

37. The cell of claim 36 further comprising an expression vector comprising a nucleic acid sequence encoding SEQ ID NO:

15.

38. The cell of claim 36, wherein the cell is a bacterial cell, insect cell or mammalian cell.

39. A method of producing an antibody or a functional fragment thereof, which antibody specifically binds to beta-amyloid 1-40, and to monomeric, oligomeric or polymeric forms of beta-amyloid 1-42, wherein said method comprises expressing in a cell the nucleic acid molecule of claim 34.

40. A nucleic acid molecule comprising a nucleotide sequence encoding a Heavy Chain Variable Region (HCVR) or a fragment thereof of an antibody which is capable of specifically binding to beta-amyloid 1-40 and to monomeric, oligomeric or polymeric forms of beta-amyloid 1-42, wherein said HCVR comprises the amino acid sequence of SEQ ID NO:

15.

41. An expression vector comprising the nucleic acid molecule of claim 40.

42. A cell comprising the expression vector of claim 41.

43. The cell of claim 42, wherein the cell is a bacterial cell, insect cell or mammalian cell.

44. A method of producing an antibody or a functional fragment thereof, which antibody specifically binds to beta-amyloid 1-40, and to monomeric, oligomeric or polymeric forms of beta-amyloid 1-42, wherein said method comprises expressing in a cell the nucleic acid molecule of claim 40.

45. A nucleic acid molecule comprising a nucleotide sequence encoding an antibody or a fragment thereof which antibody or fragment thereof is capable of specifically binding to beta-amyloid peptide 1-40 and to monomeric, oligomeric or polymeric forms of beta-amyloid 1-42, wherein said antibody comprises a Light Chain Variable Region (LCVR) comprising the amino acid of SEQ ID NO:

12 and a Heavy Chain Variable Region (HCVR) comprising the amino acid sequence of SEQ ID NO;

15.

46. A nucleic acid molecule comprising a nucleotide sequence encoding an antibody or a fragment thereof which antibody or fragment thereof is capable of specifically binding to beta-amyloid 1-40 and to monomeric, oligomeric or polymeric forms of beta-amyloid 1-42, wherein said antibody or fragment thereof comprises:

(i) human- or primate-derived framework regions,

(ii) an LCVR CDR1 comprising the amino acid sequence of SEQ ID NO;

4,

(iii) an LCVR CDR2 comprising the amino acid sequence of SEQ ID NO;

5 or the amino acid sequence of SEQ ID NO;

5 with one amino acid substitution,

(iv) an LCVR CDR3 comprising the amino acid sequence of SEQ ID NO;

6, and

(v) an HCVR comprising the amino acid sequence of SEQ ID NO;

15.

47. A nucleic acid molecule comprising a nucleotide sequence encoding an antibody or a fragment thereof which antibody or fragment thereof is capable of specifically binding to beta-amyloid 1-40 and to monomeric, oligomeric or polymeric forms of beta-amyloid 1-42, wherein said antibody or fragment thereof comprises:

(i) human- or primate-derived framework regions,

(ii) an HCVR CDR1 comprising the amino acid sequence of SEQ ID NO;

1,

(iii) an HCVR CDR2 comprising the amino acid sequence of SEQ ID NO;

2,

(iv) an HCVR CDR3 comprising the amino acid sequence of SEQ ID NO;

3, and

(v) a LCVR comprising the amino acid sequence of SEQ ID NO;

12.