Multifunctional polypeptides comprising a binding site to an epitope of the nkg2d receptor complex

• US 20040038339A1
• Filed: 01/07/2003
• Published: 02/26/2004
• Est. Priority Date: 03/24/2000
• Status: Abandoned Application

• Current Assignee: Micromet AG (Amgen Inc.)
• Original Assignee: Micromet AG (Amgen Inc.)
• Inventors: Borschert, Katrin, Rietmuller, Gert, Mayer, Monika, Kufer, Peter, Kischel, Roman, Lutterbuse, Ralf, Hofmeister, Robert
• Application Number:
  US10/239,656
• Publication Number:
  US 20040038339A1
• Time in Patent Office: Days
• US Class Current: 435/69.1
• CPC Class Codes: A61K 2039/53 DNA (RNA) vaccination
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 13/08 of the prostate
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• 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 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/10 Antimycotics
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 33/02 Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis
• CPC Class Codes: A61P 33/10 Anthelmintics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 : Immunomodulators
• CPC Class Codes: A61P 37/06 : Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 37/08 : Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 5/14 : of the thyroid hormones, e.g. T3, T4
• CPC Class Codes: C07K 16/2851 : against the lectin superfamily, e.g. CD23, CD72
• CPC Class Codes: C07K 2317/31 : multispecific
• CPC Class Codes: C07K 2317/34 : Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/622 : Single chain antibody (scFv)
• CPC Class Codes: C07K 2319/00 : Fusion polypeptide

Abstract

The present invention relates to a multifunctional polypeptide comprising a first domain comprising a binding site specifically recognizing an extracellular epitope of the NKG2D receptor complex and a second domain having receptor or ligand function. Furthermore, the present invention relates to polynucleotides encoding the multifunctional polypeptide, to vectors comprising said polypeptides and to cells comprising said polypeptides or said vectors. The invention also relates to compositions comprising either of the above recited molecules, alone or in combination, as well as to specific medical uses of the multifunctional polypeptide of the invention.

Claims

1. A multifunctional polypeptide comprising

(a) a first domain comprising a binding site specifically recognizing an extracellular epitope of the NKG2D receptor complex;

and

(b) a second domain having receptor or ligand function.

2. The multifunctional polypeptide of claim 1 wherein said binding site is the binding site of an immunoglobulin chain.

3. The multifunctional polypeptide of claim 1 wherein said binding site is a natural NKG2D-ligand of said receptor complex.

4. The multifunctional polypeptide of claim 3 wherein said natural NKG2ligand is selected from the group consisting of MIC-A, MIC-B, ULBP-1 and ULBP-2.

5. The multifunctional polypeptide of claims 1 to 4 wherein said binding site specifically recognizes an extracellular epitope of NKG2D or of DAP10.

6. The multifunctional polypeptide of any one of claims 1 to 5 wherein said receptor or ligand function is an antigen binding site of antibodies or fragments or derivatives thereof against (i) tumor associated antigens, (ii) antigens of infective agents or (iii) surface markers of sub-populations of cells such as differentiation antigens (CD antigens), natural ligands or receptors or fragments thereof or modifications thereof that interact with said tumor associated antigens or surface markers, preferably (i) heregulins, binding to the tumor associated antigens erbB-2,-3 and -4, (ii) CD4 that interacts with gp 120 of HIV infected cells or (iii) melanocyte stimulating hormone (MSH) that binds to the MSH receptor on melanocytes and tumors derived therefrom (maligned melanomes) or chemokines binding to corresponding chemokine receptors, or MHC molecules or fragments thereof complexed with peptides that bind to T-cell receptors of predefined specificity and thus recognize certain T-cell sub-populations or antigen binding sites of T-cell receptors that specifically interact with predefined MHC peptide complexes or NKp46 which interacts with haemagglutinin (HA) of influenza virus.

7. The multifunctional polypeptide of claim 6 wherein said tumor-associated antigen is selected from the group consisting of Lewis Y, Muc-1, erbB-2,-3 and -4, Ep-CAM, EGF-receptor (e.g. EGFR type I or EGFR type II), EGFR deletion neoepitope, CA19-9, Muc-1, LeY, TF-, Tn- and sTn-antigen, TAG-72, PSMA, STEAP, Cora antigen, CD7, CD19 and CD20, CD22, CD25, Ig-α

and Ig-β

, A33 and G250, CD30, MCSP and gp100, CD44-v6, MT-MMPs, (MIS) receptor type II, carboanhydrase 9, F19-antigen, Ly6, desmoglein 4, PSCA, Wue-1, GD2 and GD3 as well as TM4SF-antigens (CD63, L6, CO-29, SAS) or the alpha and gamma subunit of the fetal type acetylcholinreceptor (AChR).

8. The multifunctional polypeptide of claim 6 wherein said surface marker for an infected cell is selected from the group consisting of viral envelope antigens, e.g. of human retroviruses (HTLV I and II, HIV1 and 2) or human herpes viruses (HSV1 and 2, CMV, EBV), haemagglutinin e.g. of influenza virus A, B or C, glycoprotein E1 and E2 of rubella virus or RGP of rabies virus.

9. The multifunctional polypeptide of any one of claims 1 to 8 which is a bi-specific antibody.

10. The multifunctional polypeptide of claim 9 wherein said multifunctional polypeptide is selected from the group consisting of a synthetic, a chimeric and a humanized antibody.

11. The multifunctional polypeptide of any one of claims 1 to 10 which is a single-chain.

12. The multifunctional polypeptide of any one of claims 1 to 10 wherein said two domains are connected by a polypeptide linker.

13. The multifunctional polypeptide of any one of claims 1 to 12 wherein said first and/or second domain mimic or correspond to a V_H and V_L region of a natural antibody.

14. The multifunctional polypeptide of any one of claims 1 to 13 wherein at least one of said domains is a single-chain fragment of the variable region of the antibody.

15. The multifunctional polypeptide of any one of claims 1 to 14 wherein said domains are ranged in the order V_LNKG2D-V_HNKG2D-V_HTA-V_L-TA or V_LTA-V_HTA-V_HNKG2D-V_LNKG2D, wherein the TA represents a target antigen.

16. The multifunctional polypeptide of claim 15 wherein said target antigen is selected from the group consisting of Lewis Y, Muc-1, erbB-2,-3 and -4, Ep-CAM, EGF-receptor (e.g. EGFR type I or EGFR type II), EGFR deletion neoepitope, CA19-9, Muc-1, LeY, TF-, Tn- - and sTn-antigen, TAG-72, PSMA, STEAP, Cora antigen, CD7, CD19 and CD20, CD22, CD25, Ig-α

and Ig-β

, A33 and G250, CD30, MCSP and gp100, CD44-v6, MT-MMPs, (MIS) receptor type II, carboanhydrase 9, F19-antigen, Ly6, desmoglein 4, PSCA, Wue-1, GD2 and GD3 as well as TM4SF-antigens (CD63, L6, CO-29, SAS) or the alpha and gamma subunit of the fetal type acetylcholinreceptor (AChR).

17. The multifunctional polypeptide of any one of claims 12 to 16 wherein said polypeptide linker comprises a plurality of glycine, serine and/or alanine residues.

18. The multifunctional polypeptide of any one of claims 12 to 17 wherein said polypeptide linker comprises a plurality of consecutive copies of an amino acid sequence.

19. The polypeptide of any one of claims 12 to 18 wherein said polypeptide linker comprises 1 to 5, 5 to 10 or 10 to 15 amino acid residues.

20. The multifunctional polypeptide of any one of claims 4 to 19 wherein said polypeptide linker comprises the amino acid sequence Gly-Gly-Gly-Gly-Ser.

21. The multifunctional polypeptide of any one of claims 1 to 20 comprising at least one further domain.

22. The multifunctional polypeptide of claim 21 wherein said further domain is linked by covalent or non-covalent bonds.

23. The multifunctional polypeptide of claims 21 or 22, wherein said at least one further domain comprises an effector molecule having a conformation suitable for biological activity, capable of sequestering an ion or selective binding to a solid support or to a preselected determinant.

24. The multifunctional polypeptide of any one of claims 21 to 23 wherein said further domain confers a co-stimulatory and/or a co-activating function.

25. The multifunctional polypeptide of claim 24 wherein said co-stimulatory function is mediated by a CD28-ligand or a CD137-ligand.

26. The multifunctional polypeptide of claim 25 wherein said CD28-ligand or CD137-ligand is B7-1 (CD80), B7-2 (CD86), an aptamer or an antibody or a functional fragment or a functional derivative thereof.

27. A polynucleotide which upon expression encodes a multifunctional polypeptide and/or functional parts of a multifunctional polypeptide of any one of claims 1 to 26.

28. A vector comprising the polynucleotide of claim 27.

29. A cell transfected with the polynucleotide of claim 27 or the vector of claim 28.

30. A method for the preparation of the multifunctional polypeptide and/or parts of the multifunctional polypeptide of any one of claims 1 to 26 comprising culturing a cell of claim 29 and isolating said multifunctional polypeptide or functional parts thereof from the culture.

31. A composition comprising the polypeptide of any one of claims 1 to 26, the polynucleotide of claim 27 or the vector of claim 28.

32. The composition of claim 31 further comprising a molecule conferring a co-stimulatory and/or co-activating function.

33. The composition of claim 31 wherein said co-stimulatory function is mediated by a CD28-ligand or a CD137-ligand.

34. The composition of claim 31 wherein said CD28-ligand or CD137-ligand is B7-1 (CD80), B7-2 (CD86), an aptamer or an antibody or a functional fragment or a functional derivative thereof.

35. The composition of any one of claims 31 to 34 which is a pharmaceutical composition optionally further comprising a pharmaceutically acceptable carrier.

36. The composition of any one of claims 31 to 35 which is a diagnostic composition optionally further comprising suitable means for detections.

37. Use of the multifunctional polypeptide of any one of claims 1 to 26, the polynucleotide of claim 27 or the vector of claim 28 for the preparation of a pharmaceutical composition for the treatment of cancer, infections and/or autoimmune conditions, cancer, i.e. maligne (solid) tumors and hematopoietic cancer forms (leukemias and lymphomas), benigne tumors such as benigne hyperplasia of the prostate gland (BPH), autonomous adenomes of the thyroid gland or of other endocrine glands or adenomas of the colon;

initial stages of the malignancies, infectious diseases, caused by viruses, bacteria, fungi, protozoa or helmints, auto immune diseases wherein the elimination of the subpopulation of immune cells is desired that causes the disease;

prevention of transplant rejection or allergies.

38. The use of claim 37 wherein said infection is a viral, a bacterial or a fungal infection, wherein said cancer is a head and neck cancer, gastric cancer, oesaphagus cancer, stomach cancer, colorectal cancer, coloncarcinomaa, cancer of liver and intrahepatic bile ducts, pancreatic cancer, lung cancer, small cell lung cancer, cancer of the larynx, breast cancer, mamma carcinoma, malignant melanoma, multiple myeloma, sarcomas, rhabdomyosarcoma, lymphomas, folicular non-Hodgkin-lymphoma, leukemias, T- and B-cell-leukemias, Hodgkin-lymphoma, B-cell lymphoma, ovarian cancer, cancer of the uterus, cervical cancer, prostate cancer, genital cancer, renal cancer, cancer of the testis, thyroid cancer, bladder cancer, plasmacytoma or brain cancer or wherein said autoimmune condition is ankylosing spondylitis, acute anterior uveitis, Goodpasture'"'"'s syndrome, Multiple sclerosis, Graves'"'"' disease, Myasthenia gravis, Systemic lupus erythematosus, Insulin-dependent diabetes mellitus, Rheumatoid arthritis, Pemphigus vulgaris, Hashimoto'"'"'s thyroiditis or autoimmune Hepatitis.

39. Use of the polynucleotide of claim 27 or the vector of claim 28 for the preparation of a composition for gene therapy.

40. A method for the treatment of cancer, infections or autoimmune conditions comprising introducing the polypeptide of any one of claims 1 to 26, the polynucleotide of claim 27 or the vector of claim 28 or the composition of claim 35 into a mammal affected by said malignancies or diseases.

41. A method for delaying a pathological condition comprising introducing the polypeptide of any one of claims 1 to 26, the polynucleotide of claim 27 or the vector of claim 28 or the composition of claim 35 into a mammal affected by said pathological condition.

42. The method of claim 40 or 41 wherein said mammal is a human.

43. A kit comprising the multifunctional polypeptide of any one of claims 1 to 26, the polynucleotide of claim 27, the vector of claim 28, the cell of claim 29 or the composition of any one of claims 31 to 36.