TARGETING ABCB5 FOR CANCER THERAPY

• US 20090117117A1
• Filed: 04/11/2008
• Published: 05/07/2009
• Est. Priority Date: 04/12/2007
• Status: Active Grant

• Current Assignee: Brigham and Women's Hospital Incorporated
• Original Assignee: Brigham and Women's Hospital Incorporated
• Inventors: Sayegh, Mohamed H., Frank, Natasha Y., Frank, Markus H.
• Granted Patent: US 7,928,202 B2
• Time in Patent Office: Days
• US Class Current: 514/1.100
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61K 38/47 acting on glycosyl compounds (3.2), e.g. cellulases, lactases
• CPC Class Codes: A61K 47/6851 the antibody targeting a determinant of a tumour cell
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• 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 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/732 Antibody-dependent cellular cytotoxicity [ADCC]
• CPC Class Codes: C07K 2317/734 Complement-dependent cytotoxicity [CDC]
• CPC Class Codes: C12N 15/1135 against oncogenes or tumor suppressor genes
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/3513 Protein; Peptide
• CPC Class Codes: C12N 2320/32 Special delivery means, e.g. tissue-specific
• CPC Class Codes: C12Y 302/02022 rRNA N-glycosylase (3.2.2.22)
• CPC Class Codes: G01N 2333/705 Assays involving receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: G01N 33/574 for cancer
• CPC Class Codes: G01N 33/57492 : involving compounds localized on the membrane of tumor or cancer cells

Abstract

The invention relates to methods for treating a subject by manipulating ABCB5 on a cell as well as related products. The methods include methods of treating cancer using ABCB5 binding molecules such as antibodies and fragments thereof.

Claims

1. A method of delivering a therapeutic agent to an intracellular compartment of a cell, comprising:

contacting a cell with an isolated molecule that selectively binds to ABCB5 conjugated to a therapeutic agent in an effective amount to deliver the therapeutic agent to an intracellular compartment of the cell.

2. The method of claim 1, wherein the isolated molecule that selectively binds to ABCB5 is an isolated peptide.

3. The method of claim 2, wherein the isolated peptide is an antibody or antigen binding fragment thereof.

4. The method of claim 1, wherein the therapeutic agent is a toxin.

5. The method of claim 1, wherein the therapeutic agent is an siRNA.

6. The method of claim 1, wherein the therapeutic agent is a chemotherapeutic agent.

7. The method of claim 1, wherein the therapeutic agent is a therapeutic antibody.

8. The method of claim 1, further comprising contacting a cell with an isolated molecule that selectively binds to a surface marker selected from the group consisting of CD49e, CD133, and CD166.

9. The method of claim 1, further comprising contacting a cell with an isolated molecule that selectively binds to a surface marker selected from the group consisting of BMPR1a, TIR-1, VE-cadherin (CD144) and nestin.

10. A composition, comprising

an isolated peptide that selectively binds to ABCB5 and comprises an amino acid sequence selected from the group consisting of SEQ ID NO:

1, SEQ ID NO;

2, SEQ ID NO;

3, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, and SEQ ID NO;

8, or functionally equivalent variants thereof containing conservative substitutions, wherein the isolated peptide is not mAb 3C2-1D12.

17. A method for treating a subject, comprising

administering the composition of claim 10 to a subject having cancer in an effective amount to treat the cancer.

50. The composition of claim 10, wherein the isolated peptide comprises an amino acid sequence of a ABCB5-binding CDR3-H3 or functionally equivalent variant thereof.

51. The composition of claim 10, wherein the isolated peptide comprises an amino acid sequence of a ABCB5-binding CDR2-H2 or functionally equivalent variant thereof.

52. The composition of claim 10, wherein the isolated peptide comprises an amino acid sequence of a ABCB5-binding CDR1-H1 or functionally equivalent variant thereof.

53. The composition of claim 10, wherein the isolated peptide comprises an amino acid sequence of a ABCB5-binding CDR3-L3 or functionally equivalent variant thereof.

54. The composition of claim 10, wherein the isolated peptide comprises an amino acid sequence of a ABCB5-binding CDR2-L2 or functionally equivalent variant thereof.

55. The composition of claim 10, wherein the isolated peptide comprises an amino acid sequence of a ABCB5-binding CDR1-L1 or functionally equivalent variant thereof.

56. The composition of claim 10, wherein the isolated peptide is an isolated antibody or antibody fragment.

57. The composition of claim 56, wherein the isolated antibody or antibody fragment is an intact soluble monoclonal antibody.

58. The composition of claim 56, wherein the isolated antibody or antibody fragment is an isolated monoclonal antibody fragment selected from the group consisting of an Fab, Fab′

, F(ab′

)₂, Fv, scFv, dsFv, Fd, VH dAb, and VL dAb.

59. The composition of claim 56, wherein the isolated antibody or antibody fragment enhances chemosensitization.

60. The composition of claim 10, wherein the isolated peptide is conjugated to a detectable label.

61. The composition of claim 10, further comprising a pharmaceutically acceptable carrier.

62. The composition of claim 10, wherein the isolated peptide selectively binds to ABCB5.

63. The composition of claim 56, wherein the isolated antibody or antibody fragment is a humanized antibody.

64. The composition of claim 10, wherein the isolated peptide is a scFv.

11. A composition, comprising

an isolated peptide that selectively binds to ABCB5 and comprises an amino acid sequence selected from the group consisting of SEQ ID NO:

1, SEQ ID NO;

2, SEQ ID NO;

3, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7 and SEQ ID NO;

8, or functionally equivalent variants thereof containing conservative substitutions, wherein the isolated peptide is present in an effective amount for enhancing chemosensitization in a human subject.

12. A composition, comprising

an isolated peptide that selectively binds to ABCB5 and comprises an amino acid sequence selected from the group consisting of SEQ ID NO:

1, SEQ ID NO;

2, SEQ ID NO;

3, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7 and SEQ ID NO;

8, or functionally equivalent variants thereof containing conservative substitutions, wherein the isolated peptide is co-formulated with a therapeutic agent.

13. The composition of claim 12 wherein the isolated peptide is conjugated to the therapeutic agent.

14. The composition of claim 12 wherein the therapeutic agent is selected from the group consisting of camptothecin 9-NH2, mitoxantrone, camptothecin 7-Cl, pyrazofurin, menogaril, camptothecin 20 ester, camptothecin, amsacrine, etopside, anthrapyrazole-derivitive, temiposide, camptothecin 1′

-formyl, camptothecin 10-OH, daunorubicin, doxydoxorubicin, doxorubicin, oxanthrazoole, camptothecin 11-HOMe, zorubicin, uracil mustard, piperazinedione, hepsulfam, melphalan, bisantrene, triethylenemelamine, spiromustine, Yoshi-864, chlorambucil, piperazine mustard, hydroyurea, porfiromycin, mechlorethamine, fluorodopan, mitomycin, cytarabine (araC), dianhydrogalactitol, gemcitabine, thiotepa, N,N-dibenzyl-daunomycin, teroxirone, and aphidicolin-glycinate.

15. A kit, comprising

a container housing an isolated peptide that selectively binds to ABCB5 and comprises an amino acid sequence selected from the group consisting of SEQ ID NO:

1, SEQ ID NO;

2, SEQ ID NO;

3, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7 and SEQ ID NO;

8, or functionally equivalent variants thereof containing conservative substitutions, and instructions for administering the isolated peptide to a human subject.

16. A method for treating a subject, comprising

systemically administering an isolated peptide that selectively binds to ABCB5 and comprises an amino acid sequence selected from the group consisting of SEQ ID NO:

1, SEQ ID NO;

2, SEQ ID NO;

3, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, and SEQ ID NO;

8, or functionally equivalent variants thereof containing conservative substitutions to a subject having cancer in an effective amount to treat the cancer.

18. A method for treating a subject, comprising

administering an isolated peptide that selectively binds to ABCB5 and comprises an amino acid sequence selected from the group consisting of SEQ ID NO:

1, SEQ ID NO;

2, SEQ ID NO;

3, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7 and SEQ ID NO;

8, or functionally equivalent variants thereof containing conservative substitutions and a chemotherapeutic agent, to a subject having cancer in an effective amount to treat the cancer.

19. A method for treating a subject, comprising

systemically administering an isolated antibody or antibody fragment that selectively binds to ABCB5 and a chemotherapeutic agent to a subject having cancer in an effective amount to treat the cancer.

20. An isolated peptide comprising an immunoglobulin heavy chain variable domain, wherein:

(i) CDR1-H1 comprises an amino acid sequence of SEQ ID NO. 3;

(ii) CDR2-H2 comprises an amino acid sequence of SEQ ID NO. 4; and

(iii) a CDR3-H3 sequence, wherein the isolated peptide is not mAb 3C2-1D12.

21. The isolated peptide of claim 20, wherein the CDR3-H3 comprises an amino acid sequence of SEQ ID NO. 3.

22. The isolated peptide of claim 20, wherein the isolated peptide binds to human ABCB5.

23. The isolated peptide of claim 22, wherein the isolated peptide is an antibody.

24. The isolated peptide of claim 20 further comprising a light chain variable domain wherein CDR1-L1 comprises an amino acid sequence of SEQ ID NO. 6.

25. The isolated peptide claim 24, further comprising a CDR2-L2 that comprises an amino acid sequence of SEQ ID NO. 7.

26. The isolated peptide claim 24, further comprising a CDR3-L3 that comprises an amino acid sequence of SEQ ID NO. 8.

29. An isolated peptide comprising at least two antibody variable domains comprising:

(a) a heavy chain antibody variable domain comprising the isolated peptide of claim 20.

30. The isolated peptide of claim 29 which is a single chain Fv.

31. The isolated peptide of claim 29 which is a Fab isolated peptide.

32. The isolated peptide of claim 29 which is a fully human isolated peptide.

33. The isolated peptide of claim 29, further comprising framework regions FR1, FR2, FR3, and/or FR4 for an isolated peptide variable domain corresponding to the variant CDR1-H1, CDR2-H2, CDR3-H3, wherein the framework regions are obtained from a single polypeptide template.

34. The isolated peptide of claim 33, wherein each of the framework regions comprises an amino acid sequence corresponding to the framework region amino acid sequences of polypeptide SEQ ID NO:

1.

35. An isolated peptide according to claim 29, further comprising a dimerization domain linked to the C-terminal region of a heavy chain polypeptide variable domain.

36. The isolated peptide according to claim 35, wherein the dimerization domain comprises a leucine zipper domain or a sequence comprising at least one cysteine residue.

37. The isolated peptide of claim 35, wherein the dimerization domain comprises a hinge region from an polypeptide and leucine zipper.

38. The isolated peptide of claim 35, wherein the dimerization domain is a single cysteine.

39. The isolated peptide of claim 20, wherein the isolated peptide is a monoclonal antibody.

40. The isolated peptide of claim 20, wherein the isolated peptide is a bispecific antibody.

41. The isolated peptide of claim 20, wherein the isolated peptide is a synthetic antibody.

27. An isolated peptide comprising an immunoglobulin light chain variable domain, wherein:

(i) CDR1-L1 comprises an amino acid sequence of SEQ ID NO. 6;

(ii) CDR2-L2 comprises an amino acid sequence of SEQ ID NO. 7; and

(iii) a CDR3-L3 sequence, wherein the isolated peptide is not mAb 3C2-1D12.

28. The isolated peptide of claim 27, wherein the CDR3-L3 comprises an amino acid sequence of SEQ ID NO. 8.

42. An anti-ABCB5 antibody or antigen-binding fragment thereof, having at least one human region, wherein the anti-ABCB5 antibody or antigen binding fragment competitively inhibits binding of mAb 3C2-1D12 to ABCB5.

43. The antibody of claim 42, wherein the antigen-binding fragment is selected from the group consisting of Fab, Fab′

, F(ab′

)₂, Fv, scFv, dsFv, Fd, VH dAb, and VL dAb.

44. The antibody of claim 42, wherein the antibody or antigen-binding fragment is of immunoglobulin class IgA, IgGb 1, IgG2, IgG3, IgG4 or IgM.

45. The antibody of claim 42, wherein the antibody or antigen-binding fragment comprises a human constant region and a human variable framework region.

46. The antibody of claim 42, wherein the antigen-binding fragment is a single chain antibody.

47. The antibody of claim 46, wherein the single chain antibody is a camelid antibody.

48. A humanized antibody variable domain having a functional antigen binding region, said humanized antibody variable domain comprising non-human CDR1-H1, CDR2-H2, CDR3-H3, CDR1-L1, CD12-L2, and CDR3-L3 having at least 90% homology to CDR1-H1, CDR2-H2, CDR3-H3, CDR1-L1, CDR2-L2, and CDR3-L3 of mAb 3C2-12 incorporated into a human antibody variable domain.

49. A chimeric antibody comprising a variable domain which specifically binds to ABCB5 and a constant domain, wherein the variable domain and the constant domain are from different species.