Activatable antibodies that bind interleukin-6 receptor and methods of use thereof

Patent metadata

Field	Value
Publication number	US-9487590-B2
Application number	US-201314036973-A
Country	US
Kind code	B2
Filing date	Sep 25, 2013
Priority date	Sep 25, 2012
Publication date	Nov 8, 2016
Grant date	Nov 8, 2016

Abstract

The invention relates generally to activatable antibodies that include a masking moiety (MM), a cleavable moiety (CM), and an antibody (AB) that specifically binds to interleukin-6 receptor (IL-6R), and to methods of making and using these anti-IL-6R activatable antibodies in a variety of therapeutic, diagnostic and prophylactic indications.

First claim

What is claimed:

1. An activatable antibody that in an activated state binds Interleukin-6 Receptor (IL-6R) comprising:
an antibody or an antigen binding fragment thereof (AB) that specifically binds to the IL-6R, wherein the antibody comprises a heavy chain comprising an amino acid sequence comprising SEQ ID NO: 1 and a light chain comprising an amino acid sequence comprising SEQ ID NO: 2;
a masking moiety (MM) that inhibits the binding of the AB to the IL-6R in an uncleaved state; and
a cleavable moiety (CM) that is a polypeptide that functions as a substrate for a protease.

2. The activatable antibody of claim 1 , wherein the activatable antibody has the structural arrangement from N-terminus to C-terminus as follows in the uncleaved state:
MM-CM-AB or AB-CM-MM.

3. The activatable antibody of claim 1 , wherein the activatable antibody comprises a linking peptide between the MM and the CM.

4. The activatable antibody of claim 1 , wherein the activatable antibody comprises a linking peptide between the CM and the AB.

5. The activatable antibody of claim 1 , wherein the activatable antibody comprises a first linking peptide (LP1) and a second linking peptide (LP2), and wherein the activatable antibody has the structural arrangement from N-terminus to C-terminus as follows in the uncleaved state: MM-LP1-CM-LP2-AB or AB-LP2-CM-LP1-MM.

6. The activatable antibody of claim 5 , wherein the two linking peptides need not be identical to each other.

7. The activatable antibody of claim 5 , wherein at least one of LP1 or LP2 comprises an amino acid sequence selected from the group consisting of (GS) n , (GGS) n , (GSGGS) n (SEQ ID NO: 93) and (GGGS) n (SEQ ID NO: 94), where n is an integer of at least one.

8. The activatable antibody of claim 5 , wherein at least one of LP1 or LP2 comprises an amino acid sequence selected from the group consisting of GGSG (SEQ ID NO: 95), GGSGG (SEQ ID NO: 96), GSGSG (SEQ ID NO: 97), GSGGG (SEQ ID NO: 98), GGGSG (SEQ ID NO: 99), and GSSSG (SEQ ID NO: 100).

9. The activatable antibody of claim 5 , wherein LP1 comprises the amino acid sequence GSSGGSGGSGGSG (SEQ ID NO: 101), GSSGGSGGSGG (SEQ ID NO: 112), GSSGGSGGSGGS (SEQ ID NO: 113), GSSGGSGGSGGSGGGS (SEQ ID NO: 169), GSSGGSGGSG (SEQ ID NO: 170), or GSSGGSGGSGS (SEQ ID NO: 171).

10. The activatable antibody of claim 5 , wherein LP2 comprises the amino acid sequence GSS, GGS, GGGS (SEQ ID NO: 172), GSSGT (SEQ ID NO: 102) or GSSG (SEQ ID NO: 103).

11. The activatable antibody of claim 1 , wherein the AB has an equilibrium dissociation constant of about 100 nM or less for binding to the IL-6R.

12. The activatable antibody of claim 1 , wherein the antigen binding fragment thereof is selected from the group consisting of a Fab fragment, a F(ab′) 2 fragment, a scFv, and a scAb.

13. The activatable antibody of claim 1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 6-32, 109-111, 163-168, 181, and 182.

14. The activatable antibody of claim 1 , wherein the MM has an equilibrium dissociation constant for binding to the AB which is greater than the equilibrium dissociation constant of the AB to the IL-6R.

15. The activatable antibody of claim 1 , wherein the MM does not interfere or compete with the AB for binding to the IL-6R in a cleaved state.

16. The activatable antibody of claim 1 , wherein the MM is a polypeptide of about 2 to 40 amino acids in length.

17. The activatable antibody of claim 1 , wherein the MM polypeptide sequence is different from that of the IL-6R.

18. The activatable antibody of claim 1 , wherein the MM polypeptide sequence is no more than 50% identical to the IL6-R.

19. The activatable antibody of claim 1 , wherein the MM comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 60-62 and 80-82.

20. The activatable antibody of claim 1 , wherein the MM comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 33-59, 63-79, and 83-89.

21. The activatable antibody of claim 1 , wherein the MM comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 49, SEQ ID NO: 74, and SEQ ID NO: 78.

22. The activatable antibody of claim 1 , wherein the protease is co-localized with the IL-6R in a tissue, and wherein the protease cleaves the CM in the activatable antibody when the activatable antibody is exposed to the protease.

23. The activatable antibody of claim 1 , wherein the CM is a polypeptide of up to 15 amino acids in length.

24. The activatable antibody of claim 1 , wherein the CM is a substrate for an enzyme selected from the group consisting of a matrix metalloprotease (MMP), thrombin, a neutrophil elastase, a cysteine protease, legumain, MT-SP1, and uPA.

25. The activatable antibody of claim 1 , wherein the CM comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 90-92, 104, 105, 107, 116-128, 157-162, 162, 173, 174, and 183-193.

26. The activatable antibody of claim 1 , wherein the activatable antibody comprises a spacer, wherein the spacer is joined directly to the MM and has the structural arrangement from N-terminus to C-terminus of spacer-MM-CM-AB.

27. The activatable antibody of claim 1 comprising an agent conjugated to the AB.

28. The activatable antibody of claim 27 , wherein the agent is a therapeutic agent.

29. The activatable antibody of claim 28 , wherein the agent is an antineoplastic agent.

30. The activatable antibody of claim 27 , wherein the agent is a toxin or fragment thereof.

31. The activatable antibody of claim 27 , wherein the agent is conjugated to the AB via a linker.

32. The activatable antibody of claim 31 , wherein the linker is a cleavable linker.

33. The activatable antibody of claim 1 comprising a detectable moiety.

34. The activatable antibody of claim 33 , wherein the detectable moiety is a diagnostic agent.

35. A method of treating an inflammatory disorder comprising administering a therapeutically effective amount of the activatable antibody of claim 1 to a subject in need thereof.

Assignees

• Cytomx Therapeutics Inc

Inventors

• West James William
• Sagert Jason Gary
• Hostetter Daniel Robert
• Moore Stephen James
• Nguyen Margaret Thy Luu
• Vasiljeva Olga
• Flandez Jeanne Grace

Classifications

• A61K47/65

  Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers · CPC title

• C07K2317/56

  variable (Fv) region, i.e. VH and/or VL · CPC title

• C07K16/2866 Primary

  against receptors for cytokines, lymphokines, interferons · CPC title

• A61K2039/505

  comprising antibodies · CPC title

• C07K2317/55

  Fab or Fab' · CPC title