Nkg2a monoclonal antibodies

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to immunology. What is presented is a monoclonal antibody or a fragment thereof, which causes NK-cell induced lysis of human target cells carrying HLA-E on their surface, optionally conjugated with a detectable marker. The antibody or the fragment thereof are specific to NKG2A, have heavy and light chains, each of which contains 3 CDR and does not bind specifically: human NKG2C or NXG2E and Fc receptor. There are described: a pharmaceutical composition, a method for recovering the NK-mediated lysis of target cells, a conjugate, a set and a method for detection based on the use of the antibodies. What is disclosed is a composition for treating autoimmune and inflammatory diseases.

EFFECT: use of the invention can find application to stimulate the NK-cells leading to the lysis of dendritic cells, which contribute to the pathology of autoimmune and inflammatory diseases, that can find application in medicine.

36 cl, 4 dwg, 5 tbl, 12 ex

 

The text descriptions are given in facsimile form.

1. Monoclonal antibody or its fragment, causing lysis of NK cells cells of the target person carrying on its surface HLA-E, optionally conjugated with detektivami marker characterized by
a) specific binding to NKG2A, and the antibody or fragment is characterized by a heavy chain containing the hypervariable region 1 (CDR1), CDR2 and CDR3 of the variable segment of the heavy chain with a sequence of SED ID NQ:2 and a light chain containing the hypervariable region 1 (CDR1), CDR2 and CDR3 of the variable segment light chain sequence of SED ID NO:6;
b) the fact that considerably not associated specifically with the Fc-receptor; and
C) the fact that when binding to NKG2A on NK-cell and cause the lizirovania named NK-cell target cells of a person carrying the HLA-E on the surface of target cells, when called cell-target comes in contact with named NK-cell;
g) absence of specific binding with NKG2C or NKG2E person.

2. Monoclonal antibody or its fragment according to claim 1, further characterized by the fact that it prevents the Association of NKG2A in the HLA-E.

3. Monoclonal antibody or fragment according to claim 1, where the specified cell target is autologous cell.

4. Monoclonal antibody or its fragment according to claim 1, where the specified cell target is a cell selected from dendritic cells, cancer cells, cells infected with a virus.

5. Monoclonal antibody or fragment according to any one of claims 1 to 4, further characterized by binding to NKG2A of primates (non-human).

6. Monoclonal antibody or its fragment according to claim 5, where the binding to NKG2A on NK-cell of a Primate (non-human) named antibody induces lysis of target cells of a Primate (non-human), bearing HLA-E on the surface of target cells, under the action named NK-cells, when called cell-target comes in contact with named NK-cell.

7. Monoclonal antibody or its fragment according to claim 1, comprising the hypervariable region 1 (CDR1), CDR2 and CDR3 of the variable region of the heavy chain sequence of SEQ ID NO: 2 and the hypervariable region 1 (CDR1), CDR2 and CDR3 of the variable region of the light chain sequence of SEQ ID NO: 6.

8. Monoclonal antibody or fragment according to any one of claims 1 to 7, where named the antibody comprises an Fc region of mouse or human IgG1, which has been modified to prevent binding to Fc receptor.

9. Monoclonal antibody or fragment according to any one of claims 1 to 8, where the above mentioned antibody or ecoframe are human, chimeric or humanized.

10. Monoclonal antibody or fragment according to any one of claims 1 to 9, where named the antibody comprises a constant region of human IgG4.

11. Monoclonal antibody or fragment according to any one of claims 1 to 10, where in said CDR amino acid residues refer to each CDR in accordance with the definitions or Kabat numbering system Chotia.

12. Monoclonal antibody or fragment of claim 10, where the above mentioned antibody or its fragment is obtained on the basis of nucleic acid sequence comprising SEQ ID NO:3.

13. Monoclonal antibody or fragment of claim 10, where the above mentioned antibody or its fragment is obtained on the basis of nucleic acid sequence comprising SEQ ID NO:7.

14. Pharmaceutical composition for the recovery of lysis of NK-cells target cells, bearing on its surface HLA-E, including:
a) an effective amount monoclonalnoe antibody or its fragment according to any one of claims 1 to 13; and
b) a pharmaceutically acceptable carrier or excipient.

15. The composition according to 14, where the above composition is formulated for pharmaceutical use.

16. The composition according to item 15, additionally comprising a second therapeutic agent selected from: a therapeutic agent used in the treatment of cancer, including chemotherapeutic compound, hormone, inhibit the angiogenesis, or apoptotic agent; therapeutic agent used to treat infectious diseases, including antiviral compound, a therapeutic agent, used in other immunotherapy, such as the treatment of autoimmune diseases, inflammatory diseases, and transplant rejection; cytokine; inhibitor of cytokine, immune modulating agent; hematopoietic additional connections; growth factor; agonist activating receptor of NK cells or antagonist of inhibitory receptors of NK cells.

17. Method of recovering mediated NK cell lysis of target cells in the population, including NK-cell and called the target cell, where the named NK-cell characterized by the presence of NKG2A on its surface, and named cell-target is characterized by the presence of HLA-E on its surface, and named the method involves the step of contacting the named NK-cells with a monoclonal antibody or fragment according to any one of claims 1 to 13.

18. The method according to 17, where the named NK-cell is a human cell, called the cell-target is a cell selected from dendritic cells, cancer cells, infected with the virus cells.

19. The composition according to § 15 as a medicine.

20. Composition for the treatment of autoimmune or inflammatory diseases, prepared for farmacevticheskogo application, containing monoclonal antibody or its fragment, which is characterized by:
a) specific binding to NKG2A;
b) the fact that considerably not associated specifically with the Fc-receptor; and
C) the fact that when binding to NKG2A on NK-cell and cause the lizirovania named NK-cell target cells of a person carrying the HLA-E on the surface of target cells, when called cell-target comes in contact with named NK-cell;
g) absence of specific binding with NKG2C or NKG2E person for use in the treatment of the patient.

21. The composition according to claim 20, which is the composition by 14.

22. The composition according to claim 20, intended for combination with a second therapeutic agent selected from: immunosuppressant, a corticosteroid, an inhibitor of TNF, compounds that stimulate NCR, inhibitor inhibitory KIR receptor, an inhibitor of TGF-beta-1, inhibitor of cytokines, hematopoietic growth factor, an analgesic agent, or an anti-inflammatory agent, where named the second therapeutic agent is administered either as a separate dosage form or as part of the above composition.

23. The composition according to claim 20, where called autoimmune or inflammatory disease is selected from the group consisting of autoimmune hemolytic anemia, pernicious anemia, polyarteritis polyarthritis, systemic lupus erythematosus, granulomas is Tosa's granulomatosis, autoimmune hepatitis, diseases behceta, Crohn's disease, primary biliarnogo cirrhosis, scleroderma, ulcerative colitis, rheumatoid Segren, diabetes type I diabetes, uveitis, Graves disease, thyroiditis, myocarditis, rheumatic fever, ankylosing spondylitis, rheumatoid arthritis, glomerulonephritis, sarcoidosis, dermatomyositis, myasthenia gravis, polymyositis, Guillain-Barre syndrome, multiple sclerosis, alopecia of areata, bladderworts/kempegowda, psoriasis and vitiligo.

24. The composition according to item 15, intended for use for treating cancer in a patient, where the named cancer characterized by the presence of cancer cells expressing HLA-E on their cell surface.

25. The composition according to paragraph 24, intended for combination with a second therapeutic agent selected from anti-cancer agent or anti-emetics, where named the second therapeutic agent is administered either as a separate dosage form or as part of the above composition.

26. The composition according to § 15 for use for the treatment of viral disease in a patient, where the named viral disease characterized by the presence of infective virus cells expressing HLA-E on its cell surface.

27. The composition according to claim 20, designed to be combined with the antiviral agent, where the named PR is thepoverty agent is administered either as a separate dosage form, either as part of the above composition.

28. The composition according to § 15 to apply for the induction of tolerance to the antigen in a patient, where the specified composition is used in combination with the antigen.

29. The composition according to p for use for the treatment of autoimmune diseases or allergies.

30. The composition according to § 15 for use to improve the survival rate of hematopoietic cells in a patient.

31. The composition according to item 30, intended for use in combination with a second therapeutic agent selected from anti-cancer agent, or hematopoietic growth factor, where is called the second therapeutic agent is administered either as a separate dosage form or as part of the above composition.

32. The composition according to item 30 or 31, where the named patient suffering from leukemia.

33. Conjugate for detection of binding to NKG2A antibody, or other molecule comprising the antibody according to any one of claims 1 to 13, and the detected token.

34. Conjugate on p where specified the detected marker is selected from a radioisotope, a fluorescent dye, a member of a pair of "antigen - antibody"other than antibodies against NKG2A, member of the pair of lectin - carbohydrate", avidin, Biotin, a member of a pair of "ligand - receptor" or system member "molecular implantiruemy polymer-molecule-print".

35. Kit for detection of binding to NKG2A, including:
and conju is at on p; and
b) a material containing NKG2A.

36. The method of detection in vitro binding of the antibody with NKG2A, which includes stages:
a) contacting the conjugate in accordance with paragraph 33 with material containing NKG2A;
b) quantification of the number of detected marker associated with the named material, including NKG2A;
b) contacting the conjugate according p and called antibodies to the above-mentioned material containing NKG2A;
g) quantification of the number of detected marker associated with the named material containing NKG2A, in the presence of these antibodies;
d) comparing the number of detected material defined quantitatively in stage b)with the amount determined quantitatively on stage), to determine whether associated named antibody with material containing NKG2A.



 

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