PEPTOID LIGANDS FOR ISOLATION AND TREATMENT OF AUTOIMMUNE T-CELLS

• US 20100303835A1
• Filed: 05/28/2010
• Published: 12/02/2010
• Est. Priority Date: 05/29/2009
• Status: Abandoned Application

• Current Assignee: Opko Health, Inc., Board of Regents of the University of Texas System (University of Texas System)
• Original Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Inventors: Udugamasooriya, D. Gomika, Gocke, Anne R., Kodadek, Thomas
• Application Number:
  US12/789,711
• Publication Number:
  US 20100303835A1
• Time in Patent Office: Days
• US Class Current: 424/178.1
• CPC Class Codes: A61K 49/0008 Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure
• CPC Class Codes: A61M 37/00 Other apparatus for introducing media into the body for reproduction or fertilisation A61B17/425; apparatus for iontophoresis or cataphoresis A61N1/30; Percutany, i.e. introducing medicines into the body by diffusion through the skin salt baths A61H33/04
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: C07K 7/06 having 5 to 11 amino acids
• CPC Class Codes: G01N 33/505 involving T-cells

Abstract

The present invention provides for the identification of autoreactive T cell populations from individuals having autoimmune diseases, such as multiple sclerosis and EAE. Peptoids recognized by autoreactive T cells can be used to identify various types of autoimmune disease, and can also be used to target therapies against such populations.

Claims

1. A method of identifying a ligand that is specifically recognized by autoimmune T cells comprising:

(a) providing a first T cell population from a healthy subject, wherein said population is labeled with a first detectable label;

(b) providing a second T cell population from a subject having an autoimmune disease, wherein said population is labeled with a second detectable label;

(c) contacting said first and second T cell populations with a plurality of candidate ligands; and

(d) assessing binding of said first and second T cell populations to said candidate ligands,

wherein if said ligand binds to said second T cell population but not to said first T cell population, the said ligand is recognized by autoimmune but not healthy T cells.

2. The method of claim 1, wherein said autoimmune disease is multiple sclerosis or rheumatoid arthritis.

3. The method of claim 1, wherein said ligand is a 3-mer, a 4-mer, a 5-mer, a 6-mer, a 7-mer, an 8-mer, a 9-mer or a 10-mer.

4. The method of claim 1, wherein said first and second labels are fluorescent or chemiluminescent.

5. The method of claim 1, wherein said first and second labels are quantum dots.

6. The method of claim 1, wherein said ligand is bound to a support.

7. The method of claim 6, wherein said support is a bead, a chip, a filter, a dipstick, a membrane, a polymer matrix or a well.

8. The method of claim 7, wherein contacting comprises bringing said support into contact with said first and second T cell populations at the same time.

9. The method of claim 1, wherein said T cell population comprises CD4⁺ T cells.

10. The method of claim 1, wherein said subjects are human or murine.

11. A method of removing an autoimmune T cell from a subject suffering from an autoimmune disease comprising:

(a) providing a ligand that binds specifically to autoimmune T cells, wherein said ligand is bound to a support;

(b) contacting a T cell-containing sample from said subject with said support-bound ligand for a sufficient time to permit binding of autoimmune T cells to said support-bound ligand; and

(c) separating said support from said sample.

12. The method of claim 11, further comprising returning the sample of step (c) to said subject.

13. The method of claim 11, wherein said autoimmune disease is multiple sclerosis or rheumatoid arthritis.

14. The method of claim 11, wherein said ligand is a 3-mer, a 4-mer, a 5-mer, a6-mer, a 7-mer, an 8-mer, a 9-mer or a 10-mer.

15. The method of claim 11, wherein said support is a bead, a chip, a filter, a dipstick, a membrane, a polymer matrix or a well.

16. The method of claim 11, wherein said sample is blood, cerebrospinal fluid or semen.

17. The method of claim 16, wherein said sample is blood, and said blood is obtained from said subject, treated ex vivo, and returned to said subject.

18. The method of claim 17, wherein said blood is perfused across said support-bound ligand and returned to said subject in a closed circuit.

19. The method of claim 11, further comprising obtaining said sample from said subject.

20. The method of claim 11, wherein said subject is human or murine.

21. The method of claim 11, wherein the ligand is a peptoid as described in claims 44-63.

22. A method of killing an autoimmune T cell obtained from a subject suffering from an autoimmune disease comprising:

(a) providing a ligand that binds specifically to autoimmune T cells, wherein said ligand is conjugated to a toxin; and

(b) contacting a T cell-containing sample from said subject with said conjugate for a sufficient time to permit binding of at least one autoimmune T cell to said conjugate,

wherein said conjugate causes death of said autoimmune T cell.

23. The method of claim 22, wherein said sample is treated ex vivo, and said method further comprises returning the sample to said subject.

24. The method of claim 22, wherein said autoimmune disease is multiple sclerosis or rheumatoid arthritis.

25. The method of claim 22, wherein said ligand is a 3-mer, a 4-mer, a 5-mer, a 6-mer, a 7-mer, an 8-mer, a 9-mer or a 10-mer.

26. The method of claim 22, wherein said toxin is ricin, diphtheria toxin or cholera toxin.

27. The method of claim 22, wherein said toxin is a photo-activated toxin.

28. The method of claim 22, wherein said photo-activated toxin is ruthenium(II) tris-bipydidyl, and step (b) further comprises exposing said sample to visible light.

29. The method of claim 22, wherein said sample is blood, cerebrospinal fluid or semen.

30. The method of claim 22, further comprising obtaining said sample from said subject.

31. The method of claim 22, wherein said subject is human or murine.

32. The method of claim 22, wherein the ligand is a peptoid as described in claims 44-63.

33. A method of killing an autoimmune T cell obtained from or in a subject suffering from an autoimmune disease comprising:

(a) providing a ligand that binds specifically to autoimmune T cells, wherein said ligand is conjugated to an IgG Fc-containing molecule; and

(b) contacting an autoimmune T cell population with said conjugate for a sufficient time to permit binding of at least one autoimmune T cell to said conjugate,

wherein said conjugate recruits immune effectors to said autoimmune T cells resulting in death thereof.

34. The method of claim 33, wherein said autoimmune T cell population is treated ex vivo, and further comprising returning the sample of step (b) to said subject.

35. The method of claim 33, wherein said autoimmune disease is multiple sclerosis or rheumatoid arthritis.

36. The method of claim 33, wherein said ligand is a 3-mer, a 4-mer, a 5-mer, a 6-mer, a 7-mer, an 8-mer, a 9-mer or a 10-mer.

37. The method of claim 33, wherein said IgG Fc-containing molecule is an antibody, a single chain antibody, or a Fc fragment.

38. The method of claim 37, wherein said IgG Fc-containing molecule is an antibody or a single chain antibody, and said ligand is tethered to the antigen binding site of said antibody.

39. The method of claim 38, wherein said IgG Fc-containing molecule is an Fc fragment lacking IgG variable regions, and said peptoid is tethered to the carboxy-terminus of said Fc fragment.

40. The method of claim 33, wherein said sample is blood, cerebrospinal fluid or semen.

41. The method of claim 33, further comprising obtaining said sample from said subject.

42. The method of claim 33, wherein said subject is human or murine.

43. The method of claim 33, wherein the ligand is a peptoid as described in claims 44-63.

44. A peptoid having the formula:

45. The peptoid of claim 44, wherein n is 5.

46. The peptoid of claim 44, wherein R1 is C1-C6 alkyl unsubstituted or substituted with NH₂, OH, SH, N(C1-C6 alkyl)₂, O(C1-C6 alkyl), or S(C1-C6 alkyl);

C2-C6 alkynyl unsubstituted or substituted with NH₂, OH, SH, N(C1-C6 alkyl)₂, O(C1-C6 alkyl), or S(C1-C6 alkyl);

C2-C6 alkenyl unsubstituted or substituted with NH₂, OH, SH, N(C1-C6 alkyl)₂, O(C1-C6 alkyl), or S(C1-C6 alkyl).

47. The peptoid of claim 44, wherein R1 is C1-C6 alkyl terminally substituted with a NH₂.

48. The peptoid of claim 47, wherein R1 is 4 aminobutane.

49. The peptoid of claim 44, wherein R2 is C1-C6 alkyl unsubstituted or substituted with NH₂, OH, SH, N(C1-C6 alkyl)₂, O(C1-C6 alkyl), or S(C1-C6 alkyl);

C2-C6 alkynyl unsubstituted or substituted with NH₂, OH, SH, N(C1-C6 alkyl)₂, O(C1-C6 alkyl), or S(C1-C6 alkyl);

C2-C6 alkenyl unsubstituted or substituted with NH₂, OH, SH, N(C1-C6 alkyl)₂, O(C1-C6 alkyl), or S(C1-C6 alkyl).

50. The peptoid of claim 44, wherein R2 is C1-C6 alkyl terminally substituted with a NH₂.

51. The peptoid of claim 50, wherein R1 is 4 aminobutane.

52. The peptoid of claim 44, wherein R3 is C1-C6 alkyl, C2-C6 alkynyl, or C2-C6 alkenyl.

53. The peptoid of claim 52, wherein R3 is isobutyl.

54. The peptoid of claim 44, wherein R4 is a C1-C6 alkyl terminally substituted with a NH₂ group.

55. The peptoid of claim 54, wherein R4 is a 4 aminobutane group.

56. The peptoid of claim 44, wherein R5 is a (R)-methylbenzyl group.

57. The peptoid of claim 44, wherein R6 is a furanyl group.

58. The peptoid of claim 44, wherein R7 is C1-C6 alkyl terminally substituted with a NH2.

59. The peptoid of claim 58, wherein R7 is a 4 aminobutane group.

60. The peptoid of claim 44, wherein R8 is C1-C6 alkyl.

61. The peptoid of claim 60, wherein R8 is an isobutyl group.

62. The peptoid of claim 44, wherein R1, R2, R4, and R7 are 4-aminobutane groups;

R3 and R8 are isobutyl groups;

R5 is a (R)-methylbenzyl group; and

R6 is a furanyl group.

63. The peptoid of claim 62, wherein R8 comprises a terminal lysyl, hydroxyl, or carboxyl group.