ANTI-TRKB ANTIBODIES

• US 20110097326A1
• Filed: 01/16/2009
• Published: 04/28/2011
• Est. Priority Date: 01/17/2008
• Status: Active Grant

• Current Assignee: Novartis Ag
• Original Assignee: IRM LLC
• Inventors: Luehrsen, Kenneth R.
• Granted Patent: US 8,642,035 B2
• Time in Patent Office: Days
• US Class Current: 424/135.1
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C07K 16/2863 against receptors for growth factors, growth regulators
• CPC Class Codes: C07K 2317/55 Fab or Fab'
• 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: C07K 2317/567 Framework region [FR]
• CPC Class Codes: C07K 2317/75 Agonist effect on antigen
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto

Abstract

The present invention provides improved antibodies or antigen-binding molecules that specifically recognize and agonize the tyrosine receptor kinase B (TrkB) receptor, and methods of their use. Also provided in the invention are polynucleotides and vectors that encode such molecules and host cells that harbor the polynucleotides or vectors.

Claims

1. An antibody that binds tyrosine receptor kinase B (TrkB), wherein the antibody comprises

(a) a heavy chain variable region comprising a human heavy chain V-segment, a heavy chain complementary determining region 3 (CDR3), and a heavy chain framework region 4 (FR4), and

(b) a light chain variable region comprising a human light chain V-segment, a light chain CDR3, and a light chain FR4, wherein

i) the heavy chain CDR3 comprises the amino acid sequence V(T/V)(S/T/R/N)WFAY (SEQ ID NO:

34); and

ii) the light chain CDR3 variable region comprises the amino acid sequence (S/M)QGT(H/A)(E/V/I)PYT (SEQ ID NO;

42); and

wherein the antibody is a TrkB agonist.

2. The antibody of claim 1, wherein the heavy chain V-segment shares at least 90% sequence identity to SEQ ID NO:

16, and wherein the light chain V-segment shares at least 90% sequence identity to SEQ ID NO;

24.

3. The antibody of claim 1, wherein the heavy chain V-segment shares at least 90% sequence identity to an amino acid selected from the group consisting of SEQ ID NO:

12, SEQ ID NO;

13, SEQ ID NO;

14 and SEQ ID NO;

15, and wherein the light chain V-segment shares at least 90% sequence identity to an amino acid selected from the group consisting of SEQ ID NO;

17, SEQ ID NO;

18, SEQ ID NO;

19, SEQ ID NO;

20, SEQ ID NO;

21, SEQ ID NO;

22 and SEQ ID NO;

23.

4. The antibody of claim 1, wherein:

i) the heavy chain CDR3 comprises an amino acid sequence selected from the group consisting of SEQ ID NO;

32 and SEQ ID NO;

33; and

ii) the light chain CDR3 comprises an amino acid sequence selected from the group consisting of SEQ ID NO;

40 and SEQ ID NO;

41.

5. The antibody of claim 1, wherein the heavy chain FR4 is a human germline FR4.

6. The antibody of claim 5, wherein the heavy chain FR4 is SEQ ID NO:

50.

7. The antibody of claim 5, wherein the heavy chain J-segment comprises the human germline JH4 partial sequence WGQGTLVTVSS (SEQ ID NO:

50).

8. The antibody of claim 1, wherein the light chain FR4 is a human germline FR4.

9. The antibody of claim 8, wherein the light chain FR4 is SEQ ID NO:

61.

10. The antibody of claim 8, wherein the light chain J-segment comprises the human germline Jk2 partial sequence FGQGKLEIK (SEQ ID NO:

61).

11. The antibody of claim 1, wherein the heavy chain V-segment and the light chain V-segment each comprise a complementary determining region 1 (CDR1) and a complementary determining region 2 (CDR2);

wherein;

i) the CDR1 of the heavy chain V-segment comprises an amino acid sequence of SEQ ID NO;

27;

ii) the CDR2 of the heavy chain V-segment comprises an amino acid sequence of SEQ ID NO;

31;

iii) the CDR1 of the light chain V-segment comprises an amino acid sequence of SEQ ID NO;

38; and

iv) the CDR2 of the light chain V-segment comprises an amino acid sequence of SEQ ID NO;

39.

12. The antibody of claim 11, wherein

i) the CDR1 of the heavy chain V-segment comprises SEQ ID NO:

25;

ii) the CDR2 of the heavy chain V-segment comprises SEQ ID NO;

28;

iii) the heavy chain CDR3 comprises SEQ ID NO;

32;

iv) the CDR1 of the light chain V-segment comprises SEQ ID NO;

35;

v) the CDR2 of the light chain V-segment comprises SEQ ID NO;

38; and

vi) the light chain CDR3 comprises SEQ ID NO;

41.

13. The antibody of claim 11, wherein

i) the CDR1 of the heavy chain V-segment comprises SEQ ID NO:

25;

ii) the CDR2 of the heavy chain V-segment comprises SEQ ID NO;

28;

iii) the heavy chain CDR3 comprises SEQ ID NO;

32;

iv) the CDR1 of the light chain V-segment comprises SEQ ID NO;

36;

v) the CDR2 of the light chain V-segment comprises SEQ ID NO;

38; and

vi) the light chain CDR3 comprises SEQ ID NO;

40.

14. The antibody of claim 11, wherein

i) the CDR1 of the heavy chain V-segment comprises SEQ ID NO:

25;

ii) the CDR2 of the heavy chain V-segment comprises SEQ ID NO;

28;

iii) the heavy chain CDR3 comprises SEQ ID NO;

32;

iv) the CDR1 of the light chain V-segment comprises SEQ ID NO;

37;

v) the CDR2 of the light chain V-segment comprises SEQ ID NO;

38; and

vi) the light chain CDR3 comprises SEQ ID NO;

40.

15. The antibody of claim 11, wherein

i) the CDR1 of the heavy chain V-segment comprises SEQ ID NO:

25;

ii) the CDR2 of the heavy chain V-segment comprises SEQ ID NO;

29;

iii) the heavy chain CDR3 comprises SEQ ID NO;

32;

iv) the CDR1 of the light chain V-segment comprises SEQ ID NO;

35;

v) the CDR2 of the light chain V-segment comprises SEQ ID NO;

38; and

vi) the light chain CDR3 comprises SEQ ID NO;

41.

16. The antibody of claim 1, wherein the heavy chain variable region shares at least 90% amino acid sequence identity to the variable region of SEQ ID NO:

4 and the light chain variable region shares at least 90% amino acid sequence identity to the variable region of SEQ ID NO;

11.

17. The antibody of claim 1, wherein the heavy chain variable region shares at least 90% amino acid sequence identity to the variable region selected from the group consisting of SEQ ID NO:

1, SEQ ID NO;

2 and SEQ ID NO;

3 and the light chain variable region shares at least 90% amino acid sequence identity to the variable region selected from the group consisting of SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

8, SEQ ID NO;

9 and SEQ ID NO;

10.

18. The antibody of claim 1, wherein the heavy chain variable region shares at least 95% amino acid sequence identity to the variable region selected from the group consisting of SEQ ID NO:

1, SEQ ID NO;

2 and SEQ ID NO;

3 and the light chain variable region shares at least 95% amino acid sequence identity to the variable region selected from the group consisting of SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

8, SEQ ID NO;

9 and SEQ ID NO;

10.

19. The antibody of claim 1, wherein the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:

1, SEQ ID NO;

2 and SEQ ID NO;

3 and the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

8, SEQ ID NO;

9 and SEQ ID NO;

10.

20. The antibody of claim 1, wherein the antibody binds to TrkB with an equilibrium dissociation constant (K_D) of less than 5×

10<sup>−

8</sup> M.

21. The antibody of claim 1, wherein the antibody is a FAb′

fragment.

22. The antibody of claim 1, wherein the antibody is an IgG.

23. The antibody of claim 1, wherein the antibody is a single chain antibody (scFv).

24. The antibody of claim 1, wherein the antibody comprises human constant regions.

25. The antibody of claim 1, wherein the antibody does not bind to Tyrosine Kinase Receptor A or Tyrosine Kinase Receptor C.

26. The antibody of claim 1, wherein the antibody binds to the Ligand Binding Domain (LBD) of TrkB.

27. The antibody of claim 1, wherein the antibody competes with the binding of Brain Derived Neurotrophic Factor (BDNF) to TrkB.

28. The antibody of claim 1, wherein the antibody comprises a heavy chain comprising SEQ ID NO:

1 and a light chain comprising SEQ ID NO;

8.

29. The antibody of claim 1, wherein the antibody comprises a heavy chain comprising SEQ ID NO:

1 and a light chain comprising SEQ ID NO;

9.

30. The antibody of claim 1, wherein the antibody comprises a heavy chain comprising SEQ ID NO:

1 and a light chain comprising SEQ ID NO;

10.

31. The antibody of claim 1, wherein the antibody comprises a heavy chain comprising SEQ ID NO:

3 and a light chain comprising SEQ ID NO;

8.

33. A pharmaceutically acceptable composition comprising an antibody of claim 1 and a physiologically compatible excipient.

34. The composition of claim 33, wherein the pharmaceutical composition further comprises an agent that reduces blood glucose levels in an individual.

35. The composition of claim 33, wherein the pharmaceutical composition further comprises an agent that reduces body weight in an individual.

36. A method of reducing blood glucose levels and/or body weight in an individual in need thereof, the method comprising administering a therapeutically effective amount of the antibody of claim 1 to the individual.

37. The method of claim 36, wherein the individual is pre-diabetic.

38. The method of claim 36, wherein the individual has type I diabetes.

39. The method of claim 36, wherein the individual has type II diabetes.

40. The method of claim 36, wherein the individual is overweight.

41. The method of claim 36, wherein the individual is obese.

42. The method of claim 36, further comprising administering a therapeutically effective amount of a second agent effective in reducing blood glucose to the individual.

43. The method of claim 42, wherein the second agent and the antibody are administered as a mixture.

44. The method of claim 42, wherein the second agent and the antibody are administered separately.

45. The method of claim 42, wherein the second agent is selected from the group consisting of:

insulin, sulfonylureas, insulinotropic agents, metformin, PPARγ

agonists PPARα

agonists, PPARδ

agonists, PPARα

/γ

dual agonists, PPARα

/γ

/δ

pan agonists, alpha-glucosidase inhibitors, DPP-IV inhibitors, and GLP-1/GLP-1 analogs.

46. The method of claim 36, further comprising administering a therapeutically effective amount of a second agent effective in reducing weight or obesity is administered to the individual.

47. The method of claim 46, wherein the second agent and the antibody are administered as a mixture.

48. The method of claim 46, wherein the second agent is administered separately.

49. The method of claim 46, wherein the second agent is selected from the group consisting of lipase inhibitors, sibutramine, CB-1 inhibitors, topiramate, amylin, amylin analogs, leptin, PYY/PYY analogs, and GLP-1/GLP-1 analogs.

50. A method to reducing or inhibiting symptoms of a disorder resulting from a brain-derived neurotropic factor (BDNF) deficiency, comprising administering to a subject in need thereof a therapeutically effective amount of the antibody of claim 1 to the individual.

51. The method of claim 50, wherein the disorder resulting from a BDNF deficiency is selected from the group consisting of Rett Syndrome, Wilms'"'"' tumor, aniridia, genitourinary anomalies and mental retardation (WAGR) syndrome, obesity resulting from melanocortin receptor MC4R deficiency, and cachexia/muscle wasting syndromes.

52. The method of claim 50, wherein the disorder resulting from a BDNF deficiency is Rett Syndrome.

53. The method of claim 50, wherein the antibody is administered systemically.

32. An antibody that specifically binds TrkB, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region and the light chain variable region each comprise the following three complementary determining regions (CDRs):

CDR1, CDR2 and CDR3;

wherein;

i) the CDR1 of the heavy chain variable region comprises an amino acid sequence of SEQ ID NO;

25;

ii) the CDR2 of the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO;

28, SEQ ID NO;

29 and SEQ ID NO;

30;

iii) the CDR3 of the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO;

32 and SEQ ID NO;

33;

iv) the CDR1 of the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO;

35, SEQ ID NO;

36 and SEQ ID NO;

37;

v) the CDR2 of the light chain variable region comprises an amino acid sequence of SEQ ID NO;

39;

vi) the CDR3 of the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO;

40 and SEQ ID NO;

41.