LTBP-COMPLEX-SPECIFIC TGFβ INHIBITORS AND THEIR APPLICATION

Abstract

Inhibitors, such as antibodies and antigen-binding portions thereof, that selectively bind to LTBP1-TGFβ and / or LTBP3-TGFβ complexes are described. The invention also provides methods for using these inhibitors, for example, to inhibit TGFβ activation and to treat subjects suffering from TGFβ-associated disorders, such as fibrotic conditions. Methods for selecting a context-dependent or context-independent isoform-specific TGFβ inhibitor for a subject in need thereof are also provided.

Claims

1. An antibody or antigen-binding fragment thereof that specifically binds to the human LTBP1-TGFβ1 complex and the human LTBP3-TGFβ1 complex, comprising the following six CDRs: a) CDR-H1 comprising the amino acid sequence FTF(X1)(X2)YVMH, where X1 is S or R; and X2 is G or S; b) CDR-H2 comprising the amino acid sequence (X1)ISHEGS(X2)KYYADSVKG, where X1 is V or S; X2 is F or L; and c) CDR-H3 comprising the amino acid sequence A(X1)PRI(X2)ARRGGFGY, where X1 is R or V; X2 is A or L; d) CDR-L1 comprising the amino acid sequence TRS(X1)G(X2)ID(X3)NYVQ, wherein X1 is S or H; X2 is N, L, S or A; and X3 is N, D or Y; e) CDR-L2 comprising the amino acid sequence ED(X1)(X2)RPS, where X1 is N, F, or A; and X2 is Q, I, or V; and f) CDR-L3 comprising the amino acid sequence Q(X1)YD(X2)(X3)(X4)Q(X5)VV, where X1 is S or G; X2 is S, F, Y, D, H or W; X3 is N, D or S; X4 is N, A, L, E or T; and X5 is G, R, A or L.

2. The antibody according to claim 1, characterized in that the antibody comprises a heavy chain variable region having an amino acid sequence that is at least 90% identical to SEQ ID NO: 88; and a light chain variable region having an amino acid sequence that is at least 90% identical to SEQ ID NO:

89.

3. An antibody or its antigen-binding fragment according to any one of paragraphs 1, 2, characterized in that: a) CDR-H1 comprises the amino acid sequence of SEQ ID NO: 166; b) CDR-H2 comprises the amino acid sequence of SEQ ID NO: 167; c) CDR-H3 comprises the amino acid sequence of SEQ ID NO: 168; d) CDR-L1 comprises the amino acid sequence of SEQ ID NO: 169; e) CDR-L2 comprises the amino acid sequence of SEQ ID NO: 170; and f) CDR-L3 comprises the amino acid sequence of SEQ ID NO:

171.

4. An antibody or antigen-binding fragment thereof according to claim 3, characterized in that the antibody or antigen-binding fragment thereof comprises a heavy chain variable region having an amino acid sequence that is at least 90% identical to SEQ ID NO: 318; and a light chain variable region having an amino acid sequence that is at least 90% identical to SEQ ID NO:

319.

5. An antibody or antigen-binding fragment thereof according to claim 1, which comprises the CDR sequences: i) CDR-H1 as set forth in SEQ ID NO: 116; ii) CDR-H2 as set forth in SEQ ID NO: 117; and iii) CDR-H3 as indicated in SEQ ID NO:

118.

6. An antibody or antigen-binding fragment thereof according to any one of the preceding claims, which comprises a heavy chain variable region sequence as set forth in SEQ ID NO:

122.

7. An antibody or antigen-binding fragment thereof according to any of the preceding claims that binds the human LTBP1-proTGFβ1 complex and the human LTBP3-TGFβ1 complex with a KD of < 5 nM when measured by biolayer interferometry (BLI), optionally < 1 nM.

8. An antibody or antigen-binding fragment thereof according to any of the preceding claims, which is cross-reactive with mouse LTBP1-proTGFβ1.

9. An antibody or antigen-binding fragment thereof according to any of the preceding claims, which is cross-reactive with mouse LTBP3-proTGFβ1.

10. An antibody or antigen-binding fragment thereof according to any one of the preceding claims, wherein the antibody or antigen-binding fragment thereof binds the mouse LTBP1-proTGFβ1 complex with a KD < 10 nM when measured by biolayer interferometry (BLI).

11. An antibody or antigen-binding fragment thereof according to any one of the preceding claims, wherein the antibody or antigen-binding fragment thereof binds the mouse LTBP3-proTGFβ1 complex with a KD < 10 nM when measured by biolayer interferometry (BLI).

12. An antibody or antigen-binding fragment thereof according to any one of the preceding claims, characterized in that the antibody or antigen-binding fragment thereof cross-reacts with human and mouse LTBP1-proTGFβ1 and LTBP3-proTGFβ1 complexes, each with a KD of < 5 nM or optionally < 1 nM.

13. An antibody or antigen-binding fragment thereof according to any one of the preceding claims, characterized in that the antibody is of the IgG4 or IgG1 subtype, optionally characterized in that the antibody is of the human IgG4 subtype and comprises a substitution of Ser for Pro in the framework that forms an IgG1-like hinge.

14. A pharmaceutical composition comprising an antibody according to any of the preceding claims and a pharmaceutically acceptable carrier.

15. The pharmaceutical composition according to claim 14, which is prepared for intravenous administration or subcutaneous administration.

16. A kit for treating a TGFβ1-associated disorder, comprising a multi-dose vial containing the pharmaceutical composition of claim 14 or 15.

17. A kit for treating a TGFβ1-associated disorder, comprising a single-dose syringe containing the pharmaceutical composition of claim 14 or 15.

18. The kit according to claim 17, wherein the syringe is a disposable syringe.

19. Use of a composition according to any one of claims 14, 15 for treating a TGFβ1-associated fibrotic disorder in a human subject.

20. Use of the kit according to any one of claims 16-18 for treating a TGFβ1-associated fibrotic disorder in a human subject.

21. Use according to paragraph 19 or 20, characterized in that the fibrous disorder is fibrosis of an organ.

22. Use in accordance with paragraph 21, characterized in that the organ fibrosis is late-stage organ fibrosis.

23. Use according to paragraph 21 or 22, characterized in that the organ fibrosis is selected from the group consisting of: kidney fibrosis, liver fibrosis, pulmonary fibrosis, cardiac fibrosis, pancreatic fibrosis, skin fibrosis, scleroderma, muscle fibrosis, uterine fibrosis and endometriosis.

24. Use according to paragraph 23, characterized in that the pulmonary fibrosis is idiopathic pulmonary fibrosis (IPF).

25. Use according to paragraph 23, characterized in that the subject has chronic kidney disease (CKD).

26. Use according to paragraph 23, characterized in that liver fibrosis is associated with non-alcoholic steatohepatitis (NASH).

27. The use according to paragraph 23, characterized in that the subject has a fibrotic kidney disease selected from IgA nephropathy, focal and segmental glomerulosclerosis, crescentic glomerulonephritis, lupus nephritis and diabetic nephropathy.

28. Use according to paragraph 23, characterized in that the muscle fibrosis is associated with muscular dystrophy.

29. The use according to paragraph 28, wherein the muscular dystrophy is Duchenne muscular dystrophy (DMD).

30. Use according to any of paragraphs 19-29, wherein the antibody is administered to the subject at a dosage of 0.1 to 30 mg / kg.

31. Use according to paragraph 30, in which the antibody is administered twice a week, once a week, once every 2 weeks, once every 3 weeks, once a month or every other month.

32. Use in accordance with paragraph 30, including an initial phase of therapy and a subsequent phase of therapy, wherein the subject receives a loading dose of 2-30 mg / kg in the initial phase and then a maintenance dose of 0.1-20 mg / kg in the subsequent phase.

33. Use in accordance with paragraph 32, characterized in that the loading dose is administered to the subject twice a week or once a week.

34. Use according to paragraph 32 or 33, characterized in that the maintenance dose is administered to the subject once every 2-8 weeks.

35. Use according to paragraph 19, characterized in that the composition is used in combination with one or more additional types of therapy.

36. The use according to any of paragraphs 19-34, further comprising testing or confirming the expression of TGFβ1, LTBP1 or LTBP3 in a biological sample obtained from the patient.

37. A method for producing a pharmaceutical composition according to any one of claims 14, 15, comprising an antibody or an antigen-binding fragment thereof that specifically binds to the human LTBP1-proTGFβ complex and / or the human LTBP3-proTGFβ complex, wherein the method comprises the steps of: i) selecting an antibody or antigen-binding fragment thereof that dissociates from the human LTBP1-proTGFβ complex and / or the human LTBP3-proTGFβ complex at a t½ value of at least 45 min, and ii) mixing the antibody or antigen-binding fragment with a pharmaceutically acceptable carrier, with the provision of a composition comprising an antibody or antigen-binding fragment.

38. The method of claim 37, further comprising the step of selecting an antibody or antigen-binding fragment with an IC50 of < 5 nM when measured using cell-based assays, optionally < 2 nM.

39. The method according to claim 37 or 38, further comprising the step of confirming the efficacy of the pharmaceutical composition in an in vivo preclinical model, wherein optionally the preclinical model is a liver fibrosis model, a kidney fibrosis model, or a cardiac fibrosis model.

40. The method according to any one of paragraphs 37-39, characterized in that the method further includes the step of selecting antibodies or antigen-binding fragments that are cross-reactive with respect to human and rodent antigens.

41. The method according to any one of paragraphs 37-40, characterized in that the method further comprises the step, after step (i): subjecting the antibody or antigen-binding fragment selected in step (i) to affinity maturation and / or optimization to obtain an affinity-matured and / or optimized antibody or an affinity-matured and / or optimized antigen-binding fragment.

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