Time-staggered utilization of tumor cells in combination with intact antibodies for immunization

• US 6,994,853 B1
• Filed: 09/22/1999
• Issued: 02/07/2006
• Est. Priority Date: 09/25/1998
• Status: Expired due to Fees

• Current Assignee: Trion Pharma GmbH
• Original Assignee: Trion Pharma GmbH
• Inventors: Lindhofer, Horst, Ruf, Peter
• Primary Examiner(s): Harris, Alana M.
• Assistant Examiner(s): Holleran, Anne L.
• Application Number:
  US09/787,970
• Time in Patent Office: 2,330 Days
• Field of Search: 424/130.1, 424/136.1, 424/156.1, 424/138.1, 424/154.1, 424/155.1, 424/277.1, 424/93.7, 530/387.3, 530/388.85
• US Class Current: 424/136.1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2039/5152 Tumor cells
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 39/0011 Cancer antigens
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: C07K 16/2809 against the T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 16/30 from tumour cells
• CPC Class Codes: C07K 16/468 Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
• CPC Class Codes: C07K 2317/31 multispecific

Abstract

The invention relates to the time-staggered utilization of tumor cells in combination with intact, preferably heterologous antibodies for the immunization of humans and animals.

Claims

1. A method for inducing immunity against a tumor in a patient, comprising administering to the patient in a time-staggered manner:

(1) autologous tumor cells or allogeneic tumor cells of the same tumor type each treated to prevent their survival after reinfusion; and

(2) intact bispecific and/or trispecific antibodies having the following properties of;

(a) binding to a T cell;

(b) binding to at least one antigen on said autologous tumor cell or said allogeneic tumor cell; and

(c) binding via their Fc portion (in the case of bispecific antibodies) or via a third specificity (in the case of trispecific antibodies) to Fc receptor-positive cells,

wherein there is a time interval of 1–

48 hours between the administration of (1) and the administration of (2).

2. The method according to claim 1 wherein the administration of said tumor cells is prior to or after the administration of said antibodies.

3. The method according to claim 1 wherein the interval is 1–

24 hours.

4. The method of claim 3, wherein the interval is 1–

12 hours.

5. The method of claim 4, wherein the interval is 1–

6 hours.

6. The method of claim 5, wherein the interval is 1–

4 hours.

7. The method of claim 6, wherein the interval is 2–

4 hours.

8. The method according to claim 1 wherein the antibodies are administered in an amount of about 5–

500 μ

g in each infusion.

9. The method of claim 8, wherein the antibodies are administered in an amount of about 10–

300 μ

g.

10. The method of claim 9, wherein the antibodies are administered in an amount of about 10–

100 μ

g.

11. The method of claim 10, wherein the antibodies are administered in an amount of about 10–

50 μ

g.

12. The method of claim 8, wherein the tumor cells are administered in an amount of about 10⁷–

10⁹ cells.

13. The method of claim 12, wherein the tumor cells are administered in an amount of about 10⁸ cells.

14. The method according to claim 1 wherein said Fc receptor-positive cells have an Fcγ

receptor I, II, or III.

15. The method according to claim 14 wherein said antibodies are able to bind to monocytes, makrophages, dendritic cells, “

natural killer”

cells (NK cells) and/or activated neutrophils being Fcγ

receptor I-positive cells.

16. The method according to claim 1 wherein said antibodies are capable of inducing tumor-reactive complement-binding antibodies and therefore of inducing a humoral immune response.

17. The method according to claim 1 wherein said antibodies are selected to bind to the T cells via CD2, CD3, CD4, CD5, CD6, CD8, CD28, and/or CD44.

18. The method according to claim 1 wherein said antibodies are selected so that following their binding to the Fc receptor-positive cells the expression of CD40, CD80, CD86, ICAM-1, and/or LFA-3 being co-stimulatory antigens and/or the secretion of cytokins by the Fc receptor-positive cell is initiated or increased.

19. The method according to claim 18 wherein the antibodies are selected so that the secretion of IL-1, IL-2, IL-4, IL-6, IL-8, IL-12, INF-γ

being cytokins and/or of TNF-α

is increased.

20. The method according to claim 1 wherein said bispecific antibody is selected from the group consisting of an anti-CD3 X anti-tumor-associated antigen antibody, anti-CD4 X anti-tumor-associated antigen antibody, anti-CD5 X anti-tumor-associated antigen antibody, anti-CD6 X anti-tumor-associated antigen antibody, anti-CD8 X anti-tumor-associated antigen antibody, anti-CD2 X anti-tumor-associated antigen antibody, anti-CD28 X anti-tumor-associated antigen antibody, and anti-CD44 X anti-tumor-associated antigen antibody.

21. The method according to claim 1 wherein said bispecific antibody is selected from one or more of the following combinations of isotypes:

rat-IgG2b/mouse-IgG2a,

rat-IgG2b/mouse-IgG2b,

rat-IgG2b/mouse-IgG3,

rat-IgG2b/human-IgG1,

rat-IgG2b/human-IgG2,

rat-IgG2b/human-IgG3[oriental allotype G3m(st)=binding to protein A],

rat-IgG2b/human-IgG4,

rat-IgG2b/rat-IgG2c,

mouse-IgG2a/human-IgG3[caucasian allotypes G3m(b+g)=no binding to protein A, in the following indicated as *]

mouse-IgG2a/mouse-[VH-CH1, VL-CL]-human-IgG1-[hinge]-human-IgG3*-[CH2-CH3]

mouse-IgG2a/rat-[VH-CH1, VL-CL]-human-IgG1-[hinge]-human-IgG3*-[CH2-CH3]

mouse-IgG2a/human-[VH-CH1, VL-CL]-human-IgG1-[hinge]-human-IgG3*-[CH2-CH3]

mouse-[VH-CH1, VL-CL]-human-IgG1/rat-[VH-CH1, VL-CL]-human-IgG1-[hinge]-human-IgG3*-[CH2-CH3]

mouse-[VH-CH1, VL-CL]-human-IgG4/rat-[VH-CH1, VL-CL]-human-IgG4-[hinge]-human-IgG4[N-terminal region of CH2]-human-IgG3*[C-terminal region of CH2;

>

aa position 251]-human-IgG3*[CH3]

rat-IgG2b/mouse-[VH-CH1, VL-CL]-human-IgG1-[hinge-CH2-CH3]

rat-IgG2b/mouse-[VH-CH1, VL-CL]-human-IgG2-[hinge-CH2-CH3]

rat-IgG2b/mouse-[VH-CH1, VL-CL]-human-IgG3-[hinge-CH2-CH3, oriental allotype]

rat-IgG2b/mouse-[VH-CH1, VL-CL]-human-IgG4-[hinge-CH2-CH3]

human-IgG1/human-[VH-CH1, VL-CL]-human-IgG1-[hinge]-human-IgG3*-[CH2-CH3]

human-IgG1/rat-[VH-CH1, VL-CL]-human-IgG 1-[hinge]-human-IgG4 [N-terminal region of CH2]-human-IgG3*[C-terminal region of CH2;

>

aa position 251]-human-IgG3*[CH3]

human-IgG1/mouse-[VH-CH1, VL-CL]-human-IgG1-[hinge]-human-IgG4[N-terminal region of CH2]-human-IgG3*[C-terminal region of CH2;

>

aa position 251]-human-IgG3*[CH3]

human-IgG1/rat-[VH-CH1, VL-CL]-human-IgG1-[hinge]-human-IgG2 [N-terminal region of CH2]-human-IgG3*[C-terminal region of CH2;

>

aa position 251]-human-IgG3*[CH3]

human-IgG 1/mouse-[VH-CH1, VL-CL]-human-IgG 1-[hinge]-human-IgG2 [N-terminal region of CH2]-human-IgG3*[C-terminal region of CH2;

>

aa position 251]-human-IgG3*[CH3]

human-IgG1/rat-[VH-CH1, VL-CL]-human-IgG1-[hinge]-human-IgG3*-[CH2-CH3]

human-IgG1/mouse-[VH-CH1, VL-CL]-human-IgG1-[hinge]-human-IgG3*-[CH2-CH3]

human-IgG2/human-[VH-CH1, VL-CL]-human-IgG2-[hinge]-human-IgG3*-[CH2-CH3]

human-IgG4/human-[VH-CH1, VL-CL]-human-IgG4-[hinge]-human-IgG3*-[CH2-CH3]

human-IgG4/human-[VH-CH1, VL-CL]-human-IgG4-[hinge]-human-IgG4[N-terminal region of CH2]-human-IgG3*[C-terminal region of CH2;

>

aa position 251]-human-IgG3*[CH3]

mouse-IgG2b/rat-[VH-CH1, VL-CL]-human-IgG1-[hinge]-human-IgG3*-[CH2-CH3]

mouse-IgG2b/human-[VH-CH1, VL-CL]-human-IgG1-[hinge]-human-IgG3*-[CH2-CH3]

mouse-IgG2b/mouse-[VH-CH1, VL-CL]-human-IgG1-[hinge]-human-IgG3*-[CH2-CH3]

mouse-[VH-CH1, VL-CL]-human-IgG4/rat-[VH-CH1, VL-CL]-human-IgG4-[hinge]-human-IgG4-[CH2]-human-IgG3*-[CH3]

human-IgG1/rat-[VH-CH1, VL-CL]-human-IgG1-[hinge]-human-IgG4-[CH2]-human-IgG3*-[CH3]

human-IgG1/mouse-[VH-CH1, VL-CL]-human-IgG1-[hinge]-human-IgG4-[CH2]-human-IgG3*-[CH3]

human-IgG4/human-[VH-CH1, VL-CL]-human-IgG4-[hinge]-human-IgG4-[CH2]-human-IgG3*-[CH3].

22. The method according to claim 1 wherein said antibody is a heterologous bispecific or trispecific antibody.

23. The method of claim 22, wherein the heterologous bispecific antibody is a heterologous rat/mouse bispecific antibody.

24. The method according to claim 1 wherein the trispecific antibody comprises a T cell binding arm, a tumor cell binding arm and a third specificity for binding to Fc receptor-positive cells.

25. The method according to claim 24 wherein said trispecific antibody is selected from the group consisting of an anti-CD3 X anti-tumor-associated antigen antibody, anti-CD4 X anti-tumor-associated antigen antibody, anti-CD5 X anti-tumor-associated antigen antibody, anti-CD6 X anti-tumor-associated antigen antibody, anti-CD8 X anti-tumor-associated antigen antibody, anti-CD2 X anti-tumor-associated antigen antibody, anti-CD28 X anti-tumor-associated antigen antibody, and anti-CD44 X anti-tumor-associated antigen antibody.

26. The method according to claim 1 wherein tumor cells have been treated by irradiation or by a chemical substance.

27. The method of claim 26, wherein the irradiation is gamma irradiation.

28. The method of claim 26, wherein the irradiation has a dose of about 50 to 200 Gy.

29. The method of claim 26, wherein the chemical substance is mitomycin C.

30. The method according to claim 1 wherein said antibody binds to a surface antigen on said tumor cells, wherein said surface antigen is absent from non-tumor cells or is present in an amount insufficient for destruction of said non-tumor cells by the antibody.

31. The method according to claim 30 wherein the tumor cells are subjected to a heat pretreatment to increase the immunogenicity.

32. The method according to claim 30 wherein the surface antigen is heat shock proteins or MHC class I-related MIC molecules.

33. The method according to claim 32 wherein the heat shock proteins are HSP25, Hsp60 or Hsp70 (Hsp72) or Hsp90 proteins and the MIC molecules are MIC A or MIC B molecules.

34. The method according to claim 33 wherein the surface antigens are present in an amount of at least 100 and at the most 500,000 per tumor cell.

35. The method according to claim 34 wherein the antibody is capable of activating Fc receptor-positive cells whereby the expression of cytokins and/or co-stimulatory antigens is initiated or increased.

36. The method according to claim 1 wherein the time-staggered application of the intact bispecific and/or trispecific antibodies is performed several times.

37. The method of claim 1, wherein the tumor cells are administered in an amount of about 2×

10⁴ cells.

38. The method of claim 1, wherein the tumor cells are administered in an amount of about 5,000 cells.

39. The method of claim 1, wherein the antibodies are administered in an amount of about 1–

100 μ

g.